Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Ammonium bromide is considered not to be a sensitiser (0/20 sensitisation rate) in a guinea pig maximisation study.

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
Study period:
1997-08-04 to 1997-09-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPP 81-6 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was conducted prior to existance of OECD 429 (LLNA) testing guidelines
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: David Hall Limited, Darley Oarks, Newchurch, Staffordshire, United Kingdom
- Age at study initiation: less than one year
- Weight at study initiation: 299-321 g
- Housing: in aluminium cages (48 x 61 x 25 cm) with a grid floor (dose-range finding: 2 per cage, main study: 5 per cage)
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 20°C
- Humidity (%): mean: 67%
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

IN-LIFE DATES: From: 1997-08-04 To: 1997-09-05
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Induction:
5% test substance for intradermal injection,
55% test substance for topical application (maximum practicable concentration)

Challenge:
55% test substance for topical application (maximum practicable concentration)
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Induction:
5% test substance for intradermal injection,
55% test substance for topical application (maximum practicable concentration)

Challenge:
55% test substance for topical application (maximum practicable concentration)
No. of animals per dose:
20 in test group
10 in control group
Details on study design:
RANGE FINDING TESTS:

Two animals each were used in the dose range finding phase for the selection of appropriate concentrations for intradermal and topical induction.

Dose range finding for induction:
Dose ranging for induction was conducted via intradermal injections and topical application of formulations of the test material. For the dose range finding study, hair was clipped from an area of the scapular region (intradermal injection of 1, 2, 5 or 10% ammonium bromide solution) or of both flanks (topical induction with 25, 40, 50 or 55% ammonium bromide solution) of the animals on the day prior to administration.

Dose ranging for challenge:
Concentrations used took account of the maximum practicable concentration for administration. Three weeks prior to administration of the test material, hair was clipped from an area across the scapular region of the animals. On the following day, each animal was treated via intradermal injection.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Two
- Exposure period: day 0 intradermal injection, day 7 topical application
- Control group: 2-mercaptobenzothizole (MBT) as the positive control substance
- Site: scapula region
- Duration: Removal of test substance 48 hours after topical application with sterile distilled water.
- Concentrations: 5% test substance for intradermal injection, 55% test substance for topical application


B. CHALLENGE EXPOSURE
- No. of exposures: One
- Day(s) of challenge: day 21 topical application
- Exposure period: Removal of test substance 48 hours after topical application with sterile distilled water
- Site: scapula region
- Concentrations: 55% test substance
- Evaluation (hr after challenge): 24h, 48h after challenge; dermal reactions were also assessed 24h after intradermal and topical induction application.

OTHER:
- FCA was prepared as a 50% v/v emulsion with sterile distilled water
Positive control substance(s):
yes
Remarks:
2-mercaptobenzothizole (MBT)
Positive control results:
For positive control, animals were treated with MBT at a dose level of 75% w/v in maize oil. Following challenge, 100% of test group animals and none of the control group animals reacted positively. This demonstrates the ability of the test method to identify a mild/moderate sensitizer.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
5 % / 55 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no clinical signs
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
5 % / 55 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no clinical signs
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical signs
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no clinical signs
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
5 %/ 75 %
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
-
Remarks on result:
positive indication of skin sensitisation
Remarks:
Based on results of 6-monthly check
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
5 %/ 75 %
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
-
Remarks on result:
positive indication of skin sensitisation

Result of pilot study:

Intradermal Injection:

For the selection of appropriate concentrations for induction, intense reactions were noted at a concentration of 10% ammonium bromide. Discrete to moderate reactions were noted at concentrations of 1, 2 and 5% ammonium bromide. No reactions were seen after challenge.

Topical Application:

No reactions were noted at concentrations up to 55% ammonium bromide both for the selection of inducation and challenge concentration. Animals pretreated with Sodium Lauryl Sulphate showed dry and/or flaky and scabbed test sites after challenge application.

Overall result (main study):

No positive skin responses were noted following induction or challenge and no skin sensitising potential could be identified.

Body weight performance was considered to have been satisfactory.

Two animals /1 test group and 1 control group), were humanely killed before challenge as the condition of their scapular region had exceeded the severity limit set by the project licence governing the study. There were no other clinical signs, other than skin reactions, noted.

There were no premature deaths noted.

Table A6.1.5/01-1     Result of skin sensitisation pretest with Ammonium Bromide

Application method

Concentration
% (v/v)

Skin reaction after intracutaneous or epicutaneous application

 

Animal no. 1

Animal no. 2

 

24h

48h

72h

24h

48h

72h

 

intracutaneous

10

3

3

3

3

3

3

 

5

2

2

2

1

1

1

0

 

2

1

1

0

1

0

0

 

1

0

0

0

1

0

0

 

epicutaneous

 

Animal no. 3

Animal no. 4

 

55

0a

0a

n.d.

0

0

n.d.

 

50

0

0

n.d.

0

0

n.d.

 

40

0

0

n.d.

0

0

n.d.

 

25

0

0

n.d.

0

0

n.d.

 

0= no visible change

1= discrete or patchy erythema

2= moderate and confluent erythema

3= intense erythema and swelling

a= scabbing on test site

n.d.= not determined

Table A6.1.5/01-2   Result of intradermal induction with Ammonium Bromide (main study)

 

Group and concentration

(% ammonium bromide)

Animal number

Reaction Scores

24 hours after injection

Control

(0)

1

0

2

0

3

0

4

0

5

0

6

0

7

0

8

0

9

0

10

0

Test

(5)

1

0

2

0

3

0

4

0

5

0

6

0

7

0

8

0

9

0

10

0

11

0

12

0

13

0

14

0

15

0

16

0

17

0

18

0

19

0

20

0

Table A6.1.5/01-3     Result of topical induction with Ammonium Bromide (main study)

 

Group and concentration

(% ammonium bromide)

Animal number

Reaction Scores

24 hours after injection

Control

(0)

1

0

2

0

3

0

4

0

5

0

6

0

7

0

8

0

9

0

10

0

Test

(55)

1

0

2

0

3

0

4

0

5

0

6

0

7

0

8

0

9

0

10

0

11

0

12

0

13

0

14

0

15

0

16

0

17

0

18

0

19

0

20

0

Table A6.1.5/01-4     Result of challenge with Ammonium Bromide (main study)

 

group

Animal number

Concentration ammonium bromide (%)/

Time after patch removal

Concentration

(% ammonium bromide)/response

Concentration

(% ammonium bromide)/ % positive

55

0

24h

48h

24h

48h

55

0

55

0

control

1

0

0

0

0

-

-

0

0

2

0

0

0

0

-

-

3

0

0

0

0

-

-

4

0

0

0

0

-

-

5

0

0

0

0

-

-

6

0

0

0

0

-

-

7

0

0

0

0

-

-

8

0

0

0

0

-

-

9

0

0

0

0

-

-

10

a

a

a

a

a

a

test

1

0

0

0

0

-

-

0

0

2

0

0

0

0

-

-

3

0

0

0

0

-

-

4

0

0

0

0

-

-

5

0

0

0

0

-

-

6

0

0

0

0

-

-

7

0

0

0

0

-

-

8

0

0

0

0

-

-

9

0

0

0

0

-

-

10

0

0

0

0

-

-

11

0

0

0

0

-

-

12

0

0

0

0

-

-

13

0

0

0

0

-

-

14

0

0

0

0

-

-

15

a

a

a

a

a

a

16

0

0

0

0

-

-

17

0

0

0

0

-

-

18

0

0

0

0

-

-

19

0

0

0

0

-

-

20

0

0

0

0

-

-

 

-             Negative

a             Animal dead before assessment

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the study, ammonium bromide is considered not to be a sensitizer (0/20 sensitisation rate) in guinea pigs. In accordance with CLP Regulation (EC) No 1272/2008, no classification and labelling with respect to skin sensitisation is required.
Executive summary:

Materials and Methods

The objective of the study was to determine the delayed contact hypersensitivity of ammonium bromide using the protocol of the Magnusson-Kligman Maximisation Test in guinea pigs. The induction procedure for the test group of 20 guinea pigs consisted of exposure to the test material via two routes, intradermal injection (5% ammonium bromide) and topical application (55% ammonium bromide; the maximum practicable concentration). Animals were also exposed to an adjuvant via intradermal injection. The control group was exposed to vehicle, sterile distilled water, only. Prior to a topical challenge application with 55% ammonium bromide, all animals were treated with a 10% SLS solution the day before. The appropriate concentrations for intradermal injection and topical induction as well as for the topical challenge application were selected in a dose range finding study.

Results and Discussion

Prior to challenge, 2 animals (1 test group, 1 control group) were humanely killed as the condition of their scapular region had exceeded the severity limit set by the project licence governing the study. Following challenge no positive responses were observed in any of the remaining control or test group animals after challenge with 55% ammonium bromide.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1988
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
EPA OPP 81-6 (Skin Sensitisation)
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire, England
- Age at study initiation: Not indicated
- Weight at study initiation: 301-374g
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Induction:
7.5% w/w sodium bromide, technical grade in water (intradermal injection)
30% w/w sodium bromide, technical grade in water (topical application)

Challenge:
5% and 10% w/w sodium bromide, technical grade in water
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Induction:
7.5% w/w sodium bromide, technical grade in water (intradermal injection)
30% w/w sodium bromide, technical grade in water (topical application)

Challenge:
5% and 10% w/w sodium bromide, technical grade in water
No. of animals per dose:
20 in test group
20 in control group
Details on study design:
RANGE FINDING TESTS:
The irritancy of a range of dilutions following intradermal and topical application was investigated for the selection of suitable irritant concentrations for induction and non-irritant concentrations for challenge.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Two
- Exposure period: day 0 intradermal injection, day 7 topical application
- Site: scapular region
- Duration: 24 h for topical application
- Concentrations: 7.5% w/w sodium bromide, technical grade in water (intradermal injection), 30% w/w sodium bromide, technical grade in water (topical application)


B. CHALLENGE EXPOSURE
- No. of exposures: One
- Day(s) of challenge: day 21
- Exposure period: 24 hours
- Site: scapular region
- Concentrations: 5% and 10% w/w sodium bromide, technical grade in water
- Evaluation (hr after challenge): 24h, 48h and 72h after challenge


OTHER:
Concentration Freunds Complete Adjuvant (FCA): 50% in water
Positive control substance(s):
yes
Remarks:
Formalin
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
7.5 % / 30 %, 10 % / 5 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no clinical signs
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 7.5 % / 30 %, 10 % / 5 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no clinical signs .
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
7.5 % / 30 %, 10 % / 5 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no clinical signs
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 7.5 % / 30 %, 10 % / 5 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no clinical signs .
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no clinical signs
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no clinical signs .
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no clinical signs
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no clinical signs .

Result of pilot study:

Intradermal injection of different concentrations of sodium bromide resulted in slight to mild oedema and slight to necrotic erythema (see table 6.1.5/02-1).

Topical application of 5, 10, 20 and 30 % sodium bromide, technical grade showed localized dermal reactions in one of the animals treated with 20% of test substance. This reaction was reversible and skin appeared normal by 24 hours after treatment. All other animals did not show reactions after treatment.

Based on the results of the preliminary investigations, concentrations of 7.5% (w/w) and 30% (w/w) were selected for intradermal and topical induction and 10% and 5% (w/w) for topical challenge application.

Overall result of main study:

Sodium bromide, technical grade did not produce evidence of delayed contact hypersensitivity

Table A6.1.5/02-1          Results of pre-test with Sodium Bromide, technical grade, intradermal injection

 

Guinea-pig number

Concentration

[% w/w]

 

 

Score

 

  

1

2

 

4674/4679

10

D

E

O

8/12

N/N

2/2

8/10

N/N

2/2

 

7.5

D

E

O

8/10

2/2

1/1

8/10

1/1

1/1

 

5

D

E

O

6/8

2/2

1/1

6/6

1/1

2/2

 

2.5

D

E

O

6/6

2/2

1/1

6/6

1/1

1/1

 

1

D

E

O

4/4

1/1

1/1

4/4

1/1

1/1

 

0.5

D

E

O

4/4

1/1

1/1

4/4

1/1

1/1

 

0.25

D

E

O

4/4

1/1

1/1

4/4

1/1

1/1

 

0.1

D

E

O

4/4

1/1

1/1

4/4

1/1

1/1

 

vehicle control

(water)

D

E

    O

4/4

1/1

1/1

4/4

1/1

1/1

 

D: Diameter (mm)

E: Erythema (0-4 numerical scores)

O: Oedema (0-4 numerical scores)

N: Necrosis

1: score assessed day of injection

2: score assessed day 3 after injection

Table A6.1.5/02-2          Results of main study with Sodium Bromide, technical grade, challenge application

Treatment

Guinea-pig number

Erythema (E)

Oedema (O)

Score

Results

24 h

48 h

72 h

A

P

A

P

A

P

Freund´s

Treated control

5590

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5591

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5592

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5593

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5594

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5595

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5596

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5597

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5598

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5599

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5560

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5501

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5502

E

O

L1

0

0

0

0

0

0

0

0

0

0

0

n.d.

5503

E

O

L1

0

0

0

0

0

0

0

0

0

0

0

n.d.

5504

E

O

0

0

L1

0

0

0

0

0

0

0

0

0

n.d.

5505

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5506

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5507

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5508

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

5509

E

O

0

0

0

0

0

0

0

0

0

0

0

0

n.d.

 

Test

5510

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5511

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5512

E

O

0

0

L1

0

0

0

0

0

0

0

0

0

-

5513

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5514

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5515

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5516

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5517

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5518

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5519

E

O

0

0

L1

0

0

0

0

0

0

0

0

0

-

5520

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5521

E

O

L1

0

L1

0

0

0

0

0

0

0

0

0

-

5522

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5523

E

O

L1

0

0

0

0

0

0

0

0

0

0

0

-

5524

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5525

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5526

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5527

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5528

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

5529

E

O

0

0

0

0

0

0

0

0

0

0

0

0

-

A           Anterior site, exposed to 10% w/w sodium bromide, technical grade in distilled water

P            Posterior site, exposed to 5% w/w sodium bromide, technical grade in distilled water

L1          Localized dermal reaction, restricted to a small area of the challenge site

Results: +              positive

-                    negative

+/-         inconclusive

 

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
In this screening test performed with twenty guinea pigs, sodium bromide, technical grade did not produce evidence of delayed contact hypersensitivity. The test substance does not have to be classified and labelled with respect to skin sensitisation.
Executive summary:

Materials and Methods

The study was designed to assess skin sensitisation potential of Sodium Bromide, technical grade. The intradermal and topical irritancy of a range of dilutions of sodium bromide, technical grade was investigated to identify (a) irritant test substance concentrations suitable for the induction phase of the main study and (b) non-irritant concentrations by the topical route of administration for the challenge phase. Based on the results of this preliminary investigation concentrations of sodium bromide for the main study were selected. For the main study twenty guinea pigs were treated with the test substance intracutaneously (day 0) and epicutaneously (day 7) for induction and epicutaneously (day 21) for challenge.

Results and Discussion

The dermal reactions seen in the test animals are similar to those seen in the control animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The objective of the study was to determine the delayed contact hypersensitivity of ammonium bromide using the protocol of the Magnusson-Kligman Maximisation Test in guinea pigs. The induction procedure for the test group of 20 guinea pigs consisted of exposure to the test material via two routes, intradermal injection (5% ammonium bromide) and topical application (55% ammonium bromide; the maximum practicable concentration). Animals were also exposed to an adjuvant via intradermal injection. The control group was exposed to vehicle, sterile distilled water, only. Prior to a topical challenge application with 55% ammonium bromide, all animals were treated with a 10% SLS solution the day before. The appropriate concentrations for intradermal injection and topical induction as well as for the topical challenge application were selected in a dose range finding study.

Prior to challenge, 2 animals (1 test group, 1 control group) were humanely killed as the condition of their scapular region had exceeded the severity limit set by the project licence governing the study. Following challenge no positive responses were observed in any of the remaining control or test group animals after challenge with 55% ammonium bromide.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the experimental results of the Guinea pig maximisation test, ammonium bromide is not classified for sensitization.