Ammonium chloride

Endpoint summary

Description of key information

Oral:

In an acute oral toxicity study similar to OECD guideline 401 in rat, an LD50 of 1410 mg/kg bw was determined.

Dermal:

In an acute dermal toxicity study similar to EU method B.3 in rat, an LD50 of above 2000 mg/kg bw was determined.

Inhalation:

Not a relevant route of exposure.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

(10 animals/sex/dose)

Principles of method if other than guideline:

The study was conducted according to an internal BASF method whose principle is comparable to the OECD Guideline 401. A test group consisting of 10 animals/sex/dose was treated by single gavage with an aqueous solution of the test substance. Body weights were monitored during the 14 day observation period. The animals were observed for mortality and for clinical signs of toxicity for a period of 14 days. Decedents were subjected to necropsy. At the end of the observation period, the surviving animals were sacrificed (CO2 aphyxiation) for the purpose of necropsy. The LD50 value was estimated on the basis of the observed mortalities using the method of Finney D. J (Probit analysis, Cambridge University Press, 3 Aufl., 1971).

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

- Physical state: solid
- Analytical purity: 99.7%

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr.K.THOMAE GmbH, Biberach, Germany
- Weight at study initiation: male: 170-178 g, female: 171-187 g (within 20% of mean weight)
- Fasting period before study: The animals were given no feed for 16 hours before administration, but water was available ad libitum
- Housing: 5 animals per cage (Type VII A steel cages; FA Becker & Co., Castrop-Rauxel)
- Diet: Kliba-Labordiaet, (Klingentalmuehle AG, CH); ad libitum
- Water: tap water; ad libitum
- Acclimation period: for at least one week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20- 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

- Concentration of test substance in vehicle: 6.81, 10, 14.7, 21.5 %
- Maximum Volume applied: 10 mL/kg bw

Doses:

681, 1000, 1470, 1780, 2150 mg/kg bw

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: several times on the day of administration, at least once each workday. Check for mortality and moribund animals was performed twice daily on week days and once daily on Saturday, Sunday or on public holidays.
- Frequency of weighing: days 0, 3, 4, 7, 9, 13
- Necropsy of survivors performed: yes; before sacrifice, animals were fasted for a period of 16 hours
- Other examinations performed: clinical signs, body weight, histopathology

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 410 mg/kg bw

Based on:

dissolved

Remarks:

in water

95% CL:

>= 1 260 - <= 1 550

Remarks on result:

other: Slope factor = 1.36

Mortality:

Mortality: (out of 10 males and 10 females)
- Number of dead males at doses 2150, 1780, 1470, 1000 and 681: (after 1h) 8/7/4/0/0, (after 1d) 8/7/4/0/0, (≥2-14d) 8/7/4/0/0
- Number of dead females at doses, 2150, 1780, 1470, 1000, 681: (after 1h) 10/9/7/0/0, (after 1d) 10/9/7/3/0, (≥2-14d) 10/9/7/3/0

Clinical signs:

other: No clinical signs were manifested by animals of the lowest dose group (NOEL clinical signs = 681 mg/kg bw). Clinical signs were seen in animals dosed with 1000 mg/kg bw and above. Clinical symptoms indicated an influence on the central nervous system. Fo

Gross pathology:

Dying animals:
- General congestion: poor
- Heart: dilatation bilaterally; sporadically dilatation on the left side;
- Stomach: atonic; liquid contents; malacia of the mucosa in the glandular stomach;
- Intestines: atonic, diarrheal and mucous contents; malacia of the mucosa in several cases;

Other findings:

Histopathology (only one animal):
- Liver: fatty degeneration
- Kidney: fatty degeneration, lower nephron nephrosis

Table 1: Mortality

Dose (mg/kg bw)	Conc.	Gender	1 h	24 h	48 h	day 7	day 14
2150	21.5	male	8	8	8	8	8
		female	10	10	10	10	10
1780	17.8	male	7	7	7	7	7
		female	9	9	9	9	9
1470	14.7	male	4	4	4	4	4
		female	7	7	7	7	7
1000	10.0	male	0	0	0	0	0
		female	0	3	3	3	3
681	6.81	male	0	0	0	0	0
		female	0	0	0	0	0

Table 2: Weight (g)

Dose (mg/kg bw)	Gender	day 0	day 3/4	day 7	day 9	day 13
2150	male	178	216	248	262	-
	female	178	-	-	-	-
1780	male	171	206	236	245	-
	female	179	198	210	219	-
1470	male	170	210	241	251	-
	female	179	204	220	221	-
1000	male	177	226	246	-	283
	female	179	207	213	-	226
681	male	170	221	240	-	269
	female	171	205	210	-	218

Table 3: Clinical signs

Dose (mg/kg bw)	2150		1780		1470		1000		681
	male	female	male	female	male	female	male	female	male	female
Dyspnea	<15 m-5 h	<15 m-30 m	<15 m-5 h	<15 m-5 h	<15 m-5 h	<15 m-5 h	15 m-2 h	15 m-2 h	-	-
Apathy	<15 m-1 h	<15 m-30 m	<15 m-1 h	<15 m-1 h	<15 m-1 h	<15 m-1 h	30 m-2 h	30 m-2 h	-	-
Abnormal position	<15 m-1 h	<15 m-30 m	15 m-30 m	15 m-1 h	30 m	15 m-30 m	-	30 m	-	-
Stagger	1 h-5 h	-	<15 m-5 h	<15 m-5 h	<15 m-5 h	<15m-5 h	15 m-2 h	15 m-2 h	-	-
Atony	<15 m-30 m	<15 m-30 m	15 m-30 m	15 m-1 h	30 m	30 m	-	-	-	-
Twitching	<15 m-1 h	<15 m-30 m	15 m-1 h	15 m-1 h	30 m-1 h	15 m-1 h	-	30 m	-	-
Tonic convulsions	15 m-30 m	15 m-30 m	-	30 m	-	-	-	-	-	-
Piloerection	4 h-5 h	-	4 h-5 h	4 h-5 h	4 h-5 h	4 h-5 h	-	-	-	-
Exophthalmus	<15 m-1 h	<15 m-30 m	15 m-1 h	15 m-1 h	30 m-1 h	15 m-1 h	-	-	-	-
Poor general state	<15 m-1 h	<15 m-30 m	<15 m-1 h	<15 m-1 h	<15 m-1 h	<15 m-1 h	30 m-2 h	30 m-2 h	-	-

m: minutes

h: hour

d: day

Interpretation of results:

Category 4 based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 410 mg/kg bw

Quality of whole database:

Klimisch code 2

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 April to 13 May, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: VELAZ s.r.o, Kolec u Kladna, Czech Republic
- Weight at study initiation: males: 267 - 291 g; females: 167 - 204 g
- Fasting period before study :no
- Housing: individually in plastic cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 26 April 2010 To: 13 May 2010

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 6 x 6 cm
- % coverage: 10 % of the body surface
- Type of wrap if used: application site covered by mull and held in place by plaster (strapping)

REMOVAL OF TEST SUBSTANCE
- Washing: yes
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 2000 mg/kg body weight
- For solids, paste formed: no

Duration of exposure:

24 hours

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily observations, weighing before application and on days 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: macroscopic abnormalities at necropsy

Statistics:

no

Preliminary study:

A preliminary study was performed with 1 male and 1 female at 2000 mg/kg body weight. Slight irritation of treated skin in the female on days 2-4 after application.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

none

Clinical signs:

other: Slight irritation of treated skin in 1 female on days 2-4 after application was observed.

Gross pathology:

No macroscopic abnormalities.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Guideline and GLP conform studies.

Additional information

Oral Route of Exposure

In the key study, comparable to the OECD guideline 401 (BASF SE, 1983), groups of Wistar rats (10 rats/sex/dose) were given a single oral dose (gavage) of ammonium chloride (analytical purity 99.7 %) in water at dose levels of 681, 1000, 1470, 1780 and 2150 mg/kg bw with a subsequent observation period of 14 days. Dose dependent mortality occurred, with most of the animals dying within 1 hour after application (681 mg/kg bw: 0/20, 1000 mg/kg bw: 3/20, 1470 mg/kg bw: 11/20, 1780 mg/kg bw: 16/20 and 2150 mg/kg bw: 18/20). The LD50 calculated using Probit analysis was 1410 mg/kg bw (1260 – 1550; Slope 1.36). No clinical signs were manifested by animals of the lowest dose group (NOEL clinical signs = 681 mg/kg bw). Clinical signs were seen in animals dosed with 1000 mg/kg bw and above. Clinical symptoms indicated an influence on the central nervous system. Dyspnea, apathy, abnormal position, staggering, twitching, atony, tonic convulsions, poor general state and exophthalmus were observed from 15 min to 2 h post treatment.

Further studies in different mammalian species support the categorisation. An LD50 of 1658 mg/kg bw was determined in rat (Clothier et al., 1987). An LD50 of 1300 mg/kg bw was determined in mouse (Takasaki et al, 1990). An LDLo of 1000 mg/kg bw was determined in rat, sheep and rabbit (Boyd et al., 1946; Singer et al., 1971, Abdernalden`s Handbuch, 1935). In dog, an LDLo of 600 mg/kg bw was determined (Abdernalden`s Handbuch, 1935).

 

Dermal Route of Exposure

In an acute dermal toxicity study in Wistar rats with ammonium chloride according to the EC B.3 guideline (Agrofert, 2010), the test item was applied to the skin (36 cm²) of 5 animals (male and female) under semi occlusive conditions. After 24 h the test item was removed by cleaning the application site. No mortalities were observed. Slight irritation of treated skin was observed in 1 female on days 2 - 4 after application. No effect on body weight development and no gross macroscopic changes were observed. The LD50 was determined to be above 2000 mg/kg bw. This result is supported by a study with another ammonium ion containing compound, ammonium sulfate (CAS 7783-20-0). No mortality was observed within 14 days in 5 - 6 weeks old Wistar rats (3/sex) receiving an application of 2000 mg/kg bw ammonium sulfate under open conditions (Yamanaka, 1990). The LD50 was also greater than 2000 mg/kg bw.

 

Inhalative Route of Exposure

As indicated by particle size distribution (granulometry) of the substance, respirable particles which could reach the alveolar region of the lung are not present. Inhalation is therefore not a relevant route of exposure and no testing is required. 

Nevertheless, studies with another ammonium ion containing compound, ammonium sulfate (CAS 7783-20 -2) are considered adequate for the assessment of acute inhalation toxicity of ammonium chloride if dusts should be generated that could reach the alveolar region.

The acute inhalation toxicity of ammonium sulfate aerosols (average diameter 1 - 3 µm) is very low with 8 -h LC50 values of greater than 900 mg/m³ for guinea pigs. Rats were exposed repeatedly for 8 h/d to 1000 - 1200 mg/m³ (average diameter 2 - 3 µm) without mortality. No specific signs of toxicity were reported from these studies (Pepelko et al., 1980).

Mucociliary clearance was neither significantly affected in male rabbits that were exposed to 2 mg/m³ for one hour (mass median diameter: 0.4 µm) (Schlesinger, 1984), nor in sheep that were exposed to 1.1 mg/m³ (< 1 µm) for 20 minutes (Sackner et al., 1981) nor in rats exposed to 3.6 mg/m³ (0.4 µm) for 4 h (Phalen et al., 1980).

Pulmonary resistance was slightly increased and compliance was statistical significantly decreased in guinea pigs exposed to 0.5 - 9.5 mg/m³ for one hour (Amdur et al., 1978). Pulmonary mechanics were not altered in dogs breathing ammonium sulfate aerosol at a concentration of 4.1 mg/m³ for 4 h.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Furthermore, the test item is listed in Annex I of Directive 67/548/EEC. As a result the test substance is considered to be classified for acute oral toxicity (GHS Classification Category 4) under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.

No classification is warranted for acute dermal or inhalation toxicity, according to Regulation (EC) No 1272/2008, Annex I (CLP).