Aluminium sulphate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Reliable without restrictions. Well-presented study, with relevant measurement of chemical concentrations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Principles of method if other than guideline:

Method remarks:
A patch (Whatman # filter paper, 2 x 2 cm) including 0.4 g of test material, which was dissolved in 0.4 mL of sterile distilled water, was
applied to the abraded and intact sites on the shaved backs (4 x 4 cm) of Guinea pig. It was held in place for 6 hours with three fold gauze
patches, which were applied in place with elasticity bandage (6.4 cm width) to prevent leakage.
After 6 hours, three fold gauze patches were removed and the exposed areas washed using warm water without altering the existing response or the integrityof the epidermis.
The skin of animals was examined in accordance with the sensitization grading system and scored at 1 and 24 hours after the patches removal for
induction; and 24 and 48 hours after the patches removal for challenge.
The test of induction on the skin was performed 3 times on the day 0, 7 and 14 but the test of challenge was performed 2 weeks after the final
test of induction.

GLP compliance:

yes

Remarks:

Tested by Biotoxtech, Korea,(Test No. BO2113)

Type of study:

Buehler test

Test material

Test material information

Constituent 1

Reference substance name:

Calcium sulfate dihydrate

IUPAC Name:

Calcium sulfate dihydrate

Constituent 2

Reference substance name:

10101-41-4

EC Number:

600-148-1

Cas Number:

10101-41-4

IUPAC Name:

10101-41-4

Test material form:

solid: compact

Details on test material:

- Name of test material: Calcium sulfate, dihydrate
- Molecular formula (if other than submission substance): CaSO4•2H2O
- Molecular weight (if other than submission substance): 172.171
- Smiles notation (if other than submission substance): O.O.[O-]S(=O)(=O)[O-].[Ca+2]
- Structural formula attached as image file (if other than submission substance): see Fig.1
- Substance type:inorganic
- Physical state:solid
- Density -2.32 g/cm3)
- Solubility in water - 2.05 g/L at 20 °C
- Synonyms: Alabaster
Annaline
C.I. Pigment white 25
Gypsum
Gypsum stone
Land and plaster
Light spar
Magnesia white
Mineral white
Native calcium sulfate
Precipitated calcium sulfate
Sainite
Satin spar
Sulfuric acid, calcium(2+) salt, dihydrate
Terra alba
- Calcium sulfate, dihydrate consists of colorless, monoclinic and hygroscopic crystals

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

male

Details on test animals and environmental conditions:

Test condition : Test organism
- Sex: male
- Age of animal at study sensitization: 5 to 6 weeks old
- Weight at study sensitization: 306.4 ~ 457.9 g
- Number of test animals: 40 animals
Exposure time period: 6 hours
Concentration
- Induction: 0.4 g/0.4 mL (100 %); pre-treatment had conducted with 13 %, 25 %, 50 % and 100 % of test substance.
- Challenge: 0.4 g/0.4 ml (100 %)
Route of administration: topical with an occluded patch (whatman # 3 filter paper)
Negative control: solvent only (sterile distilled water)
Positive control: 0.1 % 1-Chloro-2,4-dinitrobenzene (solvent: 10 % Propylene Glycol)
Volume of material dosed: 0.4 g/0.4 ml
Duration of exposure for induction: 6 hours/week for 3 consecutive weeks
Duration of exposure for challenge: 6 hours once in two weeks from induction
Length of rest period: 2 weeks prior to analysis
Grading system used: OECD method (Magnusson and kligman grading scale for the evaluation of challenge patch test reactions)

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

epicutaneous, occlusive

Vehicle:

water

Induction 2

Concentration / amount:

Concentration
- Induction: 0.4 g/0.4 mL (100 %); pre-treatment had conducted with 13 %, 25 %, 50 % and 100 % of test substance.
- Challenge: 0.4 g/0.4 ml (100 %)
Route of administration: topical with an occluded patch (whatman # 3 filter paper)
Negative control: solvent only (sterile distilled water)
Positive control: 0.1 % 1-Chloro-2,4-dinitrobenzene (solvent: 10 % Propylene Glycol)
Volume of material dosed: 0.4 g/0.4 ml
Duration of exposure for induction: 6 hours/week for 3 consecutive weeks
Duration of exposure for challenge: 6 hours once in two weeks from induction
Length of rest period: 2 weeks prior to analysis
Grading system used: OECD method (Magnusson and kligman grading
scale for the evaluation of challenge patch test reactions)

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

water

Challenge 2

Concentration / amount:

Concentration
- Induction: 0.4 g/0.4 mL (100 %); pre-treatment had conducted with 13 %, 25 %, 50 % and 100 % of test substance.
- Challenge: 0.4 g/0.4 ml (100 %)
Route of administration: topical with an occluded patch (whatman # 3 filter paper)
Negative control: solvent only (sterile distilled water)
Positive control: 0.1 % 1-Chloro-2,4-dinitrobenzene (solvent: 10 % Propylene Glycol)
Volume of material dosed: 0.4 g/0.4 ml
Duration of exposure for induction: 6 hours/week for 3 consecutive weeks
Duration of exposure for challenge: 6 hours once in two weeks from induction
Length of rest period: 2 weeks prior to analysis
Grading system used: OECD method (Magnusson and kligman grading
scale for the evaluation of challenge patch test reactions)

No. of animals per dose:

40

Details on study design:

- Number of test animals: 40 animals
Exposure time period: 6 hours
Concentration
- Induction: 0.4 g/0.4 mL (100 %); pre-treatment had conducted with 13 %, 25 %, 50 % and 100 % of test substance.
- Challenge: 0.4 g/0.4 ml (100 %)
Route of administration: topical with an occluded patch (whatman # 3 filter paper)
Negative control: solvent only (sterile distilled water)
Positive control: 0.1 % 1-Chloro-2,4-dinitrobenzene (solvent: 10 % Propylene Glycol)
Volume of material dosed: 0.4 g/0.4 ml
Duration of exposure for induction: 6 hours/week for 3 consecutive weeks
Duration of exposure for challenge: 6 hours once in two weeks from induction
Length of rest period: 2 weeks prior to analysis
Grading system used: OECD method (Magnusson and kligman grading
scale for the evaluation of challenge patch test reactions)

Challenge controls:

- Challenge: 0.4 g/0.4 ml (100 %)

Positive control substance(s):

yes

Remarks:

0.1 % 1-Chloro-2,4-dinitrobenzene CAS No: 97-00-7 (solvent: 10 % Propylene Glycol CAS №: 57-55-6)

Study design: in vivo (LLNA)

Vehicle:

propylene glycol

Concentration:

Concentration
- Induction: 0.4 g/0.4 mL (100 %); pre-treatment had conducted with 13 %, 25 %, 50 % and 100 % of test substance.
- Challenge: 0.4 g/0.4 ml (100 %)
Route of administration: topical with an occluded patch (whatman # 3 filter paper)
Negative control: solvent only (sterile distilled water)
Positive control: 0.1 % 1-Chloro-2,4-dinitrobenzene (solvent: 10 % Propylene Glycol)
Volume of material dosed: 0.4 g/0.4 ml

No. of animals per dose:

40 animals

Details on study design:

- Clinical signs and mortality: Emaciation, sores, lacrimation and dyspnea were occurred sporadically in some animals that belong to the
treatment groups including the control groups. Each case of death was observed at the treatment group and positive control group on the day
16 and 9, respectively. These deaths were induced by stress, so no relationship with substance could be assumed.
- Body weight: No critical difference between negative control group and treatment group including positive control group.
- Necropsy opinion: The symptom of multifocal hepatization of lung was observed from the carcass of treatment group. The symptom of left lobe
hepatization of lung and multifocal white spot of liver were observed from the carcass of positive control group. These symptoms were induced by
stress, so no relationship with substance could be assumed.

Positive control substance(s):

other: 0.1 % 1-Chloro-2,4-dinitrobenzene CAS No: 97-00-7 (solvent: 10 % Propylene Glycol CAS №: 57-55-6)

Results and discussion

Positive control results:

- Clinical signs and mortality: Emaciation, sores, lacrimation and dyspnea were occurred sporadically in some animals that belong to the
treatment groups including the control groups. Each case of death was observed at the treatment group and positive control group on the day
16 and 9, respectively. These deaths were induced by stress, so no relationship with substance could be assumed.
- Body weight: No critical difference between negative control group and treatment group including positive control group.
- Necropsy opinion: The symptom of multifocal hepatization of lung was observed from the carcass of treatment group. The symptom of left lobe
hepatization of lung and multifocal white spot of liver were observed from the carcass of positive control group. These symptoms were induced by
stress, so no relationship with substance could be assumed.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

1.24

Group:

test chemical

Dose level:

0.4 g/0.4 mL (100 %);

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No visible change,According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs.

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 1.24. Group: test group. Dose level: 0.4 g/0.4 mL (100 %);. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No visible change,According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs..

Results 2

Reading:

2nd reading

Hours after challenge:

1.24

Group:

test chemical

Dose level:

0.4 g/0.4 mL (100 %)

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No visible change,According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs.

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 1.24. Group: test group. Dose level: 0.4 g/0.4 mL (100 %). No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No visible change,According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs..

Results 3

Reading:

rechallenge

Hours after challenge:

24.48

Group:

test chemical

Dose level:

0.4 g/0.4 mL (100 %);

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No visible change,According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs.

Remarks on result:

other: see Remark

Remarks:

Reading: rechallenge. . Hours after challenge: 24.48. Group: test group. Dose level: 0.4 g/0.4 mL (100 %);. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No visible change,According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs..

In vivo (LLNA)

Results

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see Remark

Remarks:

CDNB (in 10% PG) Animal Induct Induct- Induct- number ion 1 ion 2 ion 3 Challenge 0.1% 0.1% 0.1% 0.1% 1h 24h 1h 24h 1h 24h 24h 48h 1 0 2 2 2 2 3 2 2 2 2 2 2 3 2 3 3 3 3 1 2 2 3 2 3 3 3 4 1 2 2 2 1 2 2 2 5 2 3 2 3 2 3 3 3 6 2 2 1 2 2 3 3 3 7 1 2 2 3 2 3 3 3 8 2 2 2 3 1 3 3 3 9 2 3 2 3 1 2 2 2 10 0 1 1 2 - CDNB: 1-Chloro-2,4-Dinitrobenzene PG: Propylene Glycol - : Death Magnusson and kligman grading scale for the evaluation of challenge patch test reactions 0 - No visible change 1 - Discrete or patchy erythema 2 - Moderate & confluent erythema 3 - Intense erythema & swelling

Any other information on results incl. tables

- Clinical signs and mortality: Emaciation, sores, lacrimation and  dyspnea were occurred sporadically in some animals that belong to the  treatment groups including the control groups. Each case of death was  observed at the treatment group and positive control group on the day 16  and 9, respectively. These deaths were induced by stress, so no  relationship with substance could be assumed.
- Body weight: No critical difference between negative control group and  treatment group including positive control group.
- Necropsy opinion: The symptom of multifocal hepatization of lung was  observed from the carcass of treatment group. The symptom of left lobe  hepatization of lung and multifocal white spot of liver were observed  from the carcass of positive control group. These symptoms were induced  by stress, so no relationship with substance could be assumed.

- Skin sensitization:
All test groups except positive control group had sensitization score of    0 (No visible change) for both induction and challenge test.


- stimulation index(SI)
                                                                             Calcium sulfate, dihydrate
Animal Negative    Induct-     Induct-     Induct-           
number control     ion 1       ion 2       ion 3     Challenge   
                    0.1%        0.1%        0.1%        0.1% 
                  1h  24h     1h  24h     1h  24h     24h 48h
1        0         0   0       0   0      0    0       0    0
2        0         0   0       0   0      0    0       0    0
3        0         0   0       0   0      0    0       0    0
4        0         0   0       0   0      0    0       0    0
5        0         0   0       0   0      0    0       0    0
6        0         0   0       0   0      0    0       0    0
7        0         0   0       0   0      0    0       0    0
8        0         0   0       0   0      0    0       0    0
9        0         0   0       0   0      0    0       0    0
10       0         0   0       0   0      0    0       0    0
11                 0   0       0   0      0    0       0    0
12                 0   0       0   0      0    0       0    0
13                 0   0       0   0      0    0       0    0
14                 0   0       0   0      0    0       0    0
15                 0   0       0   0      0    0       0    0
16                 0   0       0   0      0    0       0    0
17                 0   0       0   0      0    0       0    0
18                 0   0       0   0      0    0       0    0
19                 0   0       0   0      0    0       -    
20                 0   0       0   0      0    0       0    0


                                CDNB (in 10% PG)
Animal             Induct-     Induct-     Induct- 
number             ion 1       ion 2       ion 3      Challenge
                    0.1%        0.1%        0.1%        0.1%
                   1h  24h     1h  24h     1h  24h     24h  48h
1                  0    2       2   2       2   3       2    2
2                  2    2       2   3       2   3       3    3
3                  1    2       2   3       2   3       3    3 
4                  1    2       2   2       1   2       2    2
5                  2    3       2   3       2   3       3    3
6                  2    2       1   2       2   3       3    3
7                  1    2       2   3       2   3       3    3   
8                  2    2       2   3       1   3       3    3
9                  2    3       2   3       1   2       2    2
10                 0    1       1   2       -


CDNB: 1-Chloro-2,4-Dinitrobenzene
PG: Propylene Glycol
- : Death

Magnusson and kligman grading scale for the evaluation of challenge patch  test reactions
0 - No visible change
1 - Discrete or patchy erythema
2 - Moderate & confluent erythema
3 - Intense erythema & swelling

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

All test groups except positive control group had sensitization score of 0 (No visible change) for both induction and challenge test.
According to the result, skin sensitization was not observed with the test concentrations of calcium sulfate, dihydrate on the backs of guinea pigs.