Ammonium chloride

Endpoint summary

Description of key information

In a skin irritation study in rabbits, according to the guideline of the Federal Register, Vol.43, No. 163 (Microbiological and Biochemical Assay Laboratories,1985), the test item did not show skin irritating properties.

In an eye irritation study comparable to OECD guideline 405 (BASF SE, 1968), the test item showed eye irritating properties.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Reliability adopted from OECD SIDS document - Meets generally accepted scientific standards and acceptable for assessmnet (original literature not available)

Qualifier:

according to guideline

Guideline:

other: Federal Register, vol.43, no 163

Version / remarks:

1978-08-22

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot No.of test material: 50820

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rich-Glo-farms (U.S.D.A. 74-AA-29), El Campo, Texas USA
- Housing: Individually in galvanized steel cages placed over trays.
- Diet: ad libitum, Purena Complete Rabbit Chow, Lot #197C and City of Houston
- Water: ad libitum, tap water
- Acclimation period: 21 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.2 - 24.4
- Humidity (%): 40 - 60

Type of coverage:

occlusive

Preparation of test site:

other: Shaved and abraded

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g
- Type of application: the test substance was applied to gauze pads which had been moistened with 1 mL distilled-deionized water
- Restrainers: Yes

Duration of treatment / exposure:

24 hour(s)

Observation period:

4 days

Number of animals:

6

Details on study design:

TEST SITE
- Application: Two site/animal were exposed to the test substance
- Area of exposure: 1 x 1 cm
- Type of wrap if used: a polyethylene wrapper (secured with masking tape)

REMOVAL OF TEST SUBSTANCE
- Washing: No. (Test site was brushed clean)

SCORING SYSTEM: The skin was scored according to the method of Draize (see below).

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Worst case scenario based on the results reported.

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Worst case scenario based on the results reported.

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Worst case scenario based on the results reported.

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Worst case scenario based on the results reported.

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Worst case scenario based on the results reported.

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Worst case scenario based on the results reported.

Irritation parameter:

edema score

Basis:

mean

Remarks:

of all six animals (12 test sites)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Unabraded skin (12 test sites): 7 test sites were scored with a draize score of 2 while 5 test sites had a draize score of 3, 24 hours after removal of the test patch. These changes were not observed after 48, 72, 96 hours. No edema or eschar was found at any observation time point.

Abraded skin was also scored. (These values however are not included in the final assessment of skin irritation potential of the test substance).
- abraded (12 test sites; 24 hour observation time point): Erythema score of 2 at 7 sites, and an erythema score of 3 was recorded at 5 sites. These changes were not observed after 48, 72, 96 hours. No edema or eschar was found at any observation time point.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

The test substance is applied to the conjunctival sac of one eye in 2 animals. The substance was tested as powder. The animals were observed after 10 min, 1 and 3 h on the day of treatment and up to 8 days afterwards. The eyes were not washed out after 24 hours as specified in OECD Guideline 405.

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.69 and 2.82 kg

ENVIRONMENTAL CONDITIONS: not specified

Vehicle:

unchanged (no vehicle)

Controls:

other: other eye treated with talcum

Amount / concentration applied:

Amount applied: 50 mg

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM: Initially, a BASF scoring system was used to score eye changes. These values were adapted to OECD Draize scores (see below).

TOOL USED TO ASSESS SCORE: not specified

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of both animals

Time point:

other: 24 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: no effects

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24 and 72 hours

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

mean

Remarks:

of both animals

Time point:

other: 24 and 72 hours

Score:

0.167

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24 and 72 hours

Score:

2.5

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Bleeding seen after 72 hours

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24 and 72 hours

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of both animals

Time point:

other: 24 and 72 hours

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24 and 72 hours

Score:

1.5

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of both animals

Time point:

other: 24 and 72 hours

Score:

1.25

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Slight erythema and chemosis was seen in the control eyes 1 hour and 24 hours after instillation of amorphous talcum. The effects were no longer present by the 72 hour observation time point (table 3).

Table 2: OECD Draize scores after treatment of the rabbit eye with Ammonium Chloride

Time	opacity	iritis	erythema	chemosis	Comments
1 h	2/2	0/0	2/1	2/2
24 h	2/2	0/0	3/2	1/2
48 h	-/-	-/-	-/-	-/-
72 h	2/2	0/1	2/2	1/1	bleeding in animal 1
8 d	0/0	0/0	0/0	0/0

"-": data missing.

 

Table 3: OECD Draize scores after treatment of the rabbit eye with amorphous Talcum (Control eye)

Time	opacity	iritis	erythema	chemosis	Comments
1 h	0/0	0/0	2/1	1/0	Residue in both animals
24 h	0/0	0/0	2/2	0/0
48 h	-/-	-/-	-/-	-/-
72 h	0/0	0/0	0/0	0/0
8 d	0/0	0/0	0/0	0/0

"-": data missing.

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Skin Irritation

In a primary dermal irritation study, performed pursuant to method of Federal Register, Vol. 43, No. 163, 1978 (Microbiological and Biochemical Assay Laboratories,1985), 6 New Zealand White were dermally exposed to 0.5 g ammonium chloride (reagent grade) applied occlusively on a gauze patch which had been moistened with water (1 mL). Application was occlusive onto the shaved or abraded skin (1 x 1 cm) for 24 hours. At the end of exposure, exposure sites were not washed. Animals were observed until total reversal of symptoms. Irritation was scored regularly using the OECD Draize method. 24 hour after removal of the test patch from non-abraded skin, 7 of the 12 exposed sites (2 sites/animal) scored for erythema, had a draize score of 2 while the remaining 5 sites had a draize score of 3. Erythema at these sites were no longer present after 48, 72 or 96 hours. No edema or eschar was found at any observation time point. On abraded test sites, the erythema scores recorded 24 hours after termination of exposure was 2 at 7 of 12 exposed sites, and an erythema score of 3 was recorded at 5 of 12 exposed sites. These changes were not observed after 48, 72, or 96 hours. No edema or eschar was found at any observation time point. The results from the abraded skin are given in conformity with Robust Study Summary requirements, however they are not included in the final assessment of the skin irritation potential of ammonium chloride.

In another dermal irritation study (BASF AG, 1968), 2 - 4 Vienna White rabbits were treated with the test substance for 1, 5, 15 minutes and 20 hours under occlusive conditions. Moistened test patches coated with neat (2 g) ammonium chloride (100%) were attached to the clipped dorsal skins of rabbit (ca. 2.5 x 2.5 cm). After the application durations of 1, 5 and 15 minutes, the application sites were washed with undiluted Lutrol and Lutrol (50%). The application sites after 20 hours exposure were not washed. The animals were observed for 8 days and skin changes were recorded on day 1, 2, 3 and 8. Very slight skin erythema was seen within 24 hours removal of test patch for animals exposed for 1, 5 and 15 minutes. Edema was not seen after treatment periods of 1, 5 and 15 mins. After 20 hours, average scores (24, 48 and 72 hours) recorded for erythema were 2.3, 2.0, 2.0 and 0.3 for animals 1, 2, 3 and 4. Scaling was still present in 1 animal and erythema was still present in another animal at the end of the observation period (day 8). Average edema scores were 2.0 ,0, and 0 for animals 1, 2, 3 and 4, respectively.

Eye Irritation

The eye irritation potential of the test substance was determined according to the protocols of a BASF test (BASF SE, 1968) comparable to OECD guideline 405. The substance was tested as powder. 50 mg ammonium chloride (p.a.) was applied to the conjunctival sac of one eye of 2 Vienna White Albino rabbits. The other eye was treated with talcum. Eyes were not washed after instillation. After instillation, the animals eyes were scored (OECD Draize method) after 10 min, 1 and 3h on the day of treatment, then after 24 and 72 hours and 8 days after instillation. Mean draize scores after 24 and 72 hours were calculated per animal for corneal opacity, iritis, conjunctivae redness and chemosis and used for assessment of irritation potential. Redness of the mucous membrane (score: 2.5 and 2.0 for animal 1 and 2, respectively), corneal opacity (score: 2.0 and 2.0 for animal 1 and 2, respectively), chemosis (score: 1.5 and 1.0 for animal 1 and 2, respectively), and mild iritis (score: 0 and 0.33 for animal 1 and 2, respectively), were observed. These effects were fully reversible within 7 days.

 

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008 (CLP). As a result the test substance is considered not to be classified for skin irritation (UN GHS No Category) under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.

In contrast, the test substance is considered to be classified for eye irritation (UN GHS Category 2) under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.