Ammonium acetate

Endpoint summary

Description of key information

Acute oral toxicity: Weight of evidence: The LD50 was higher than 2000 mg/kg bw (2333.28-3546.59 mg/kg bw). Read-across approach from experimental results obtained with analogues Potassium Acetate and Ammonium Sulphate. The  read-across approach acute toxicity effect is lower than the subacute toxicity data obtained from an experimental study. The difference can be explained as an estimation error of the read across (the choosen value is the lowest one). Both results are higher than 2000 mg/kg/bw and  shows no oral toxicity of ammonium acetate.

Acute dermal toxicity: Weight of evidence: The LD50 was greater than 2000 mg/kg bw (> 2333.28-26556.42 mg/kg bw). Read-across approach from experimental results obtained with analogues Fumaric Acid and Ammonium Sulphate.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No specific test guideline was reported; however, a scientifically defensible approach was used to conduct the study.

Principles of method if other than guideline:

Single oral toxicity is estimated by the gastric intubation of groups of five non-fasted, Carworth-Wistar rats, four to five weeks of age and 90 to 120 grams in weight, which have been reared in the laboratory own colony and mantained from time of weaning on Rockland rat diet, complete. Dose: 3.25g/kg (2.48-4.26). The observation period was 14 days.

GLP compliance:

no

Test type:

other: method described

Limit test:

no

Species:

rat

Strain:

other: Carworth-Wistar

Sex:

male

Route of administration:

oral: gavage

Vehicle:

not specified

Doses:

3.25 g/kg bw (2.48-4.26)

No. of animals per sex per dose:

Five male rats per group

Control animals:

not specified

Sex:

male

Dose descriptor:

LD50

Effect level:

ca. 3.25 other: g/kg bw

Based on:

test mat.

Potassium acetate LD50 is 3.25 g/kg bw (2.48-4.26).

Interpretation of results:

GHS criteria not met

Conclusions:

Potassium acetate LD50 is 3.25 g/kg bw (2.48-4.26).

Executive summary:

Potassium acetate LD₅₀is 3.25 g/kg bw (2.48-4.26).

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

not specified

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Peer reviewed handbook: Sax's Dangerous Properties of Industrial Materials.

Principles of method if other than guideline:

No data provided on the method.

GLP compliance:

not specified

Test type:

other: No data

Species:

rat

Strain:

not specified

Sex:

not specified

Route of administration:

other: oral

Vehicle:

not specified

Doses:

3250 mg/kg bw

No. of animals per sex per dose:

No data

Control animals:

not specified

Sex:

not specified

Dose descriptor:

LD50

Effect level:

3 250 mg/kg bw

Based on:

test mat.

Potassium acetate oral, LD50 = 3250 mg/kg bw.

Interpretation of results:

GHS criteria not met

Conclusions:

Potassium acetate oral, LD50 = 3250 mg/kg bw.

Executive summary:

Potassium acetate oral, LD₅₀ = 3250 mg/kg bw.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Potassium Acetate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the acute oral toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (non-standard method) on the analogue Potassium Acetate.

GLP compliance:

no

Test type:

other: read-across from a non-standard study with an analogue

Sex:

male

Dose descriptor:

LD50

Effect level:

2.55 other: g/kg bw

Based on:

test mat.

95% CL:

1.95 - 3.35

Other findings:

Based on the experimental results obtained with the analogue Potassium Acetate (LD 50 for male rats = 3.25 (2.48-4.26) g/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for substance Ammonium Acetate is calculated to be 2.55 (1.95-3.35) g/kg bw.

The analogue Potassium Acetate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties. These properties are:
- a low log Pow value which is -3.72 for Potassium Acetate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 2.5 g/mL at 20 ºC for Potassium Acetate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 98.14 for Potassium Acetate and 77.08 for Ammonium Acetate.

The LD50 for substance Ammonium Acetate is calculated to be 2.55 (1.95-3.35) g/kg bw.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for substance Ammonium Acetate is calculated to be 2.55 (1.95-3.35) g/kg bw.

Executive summary:

Based on the experimental results obtained (reported under the endpoint record 07.02.01_01 KAc) with the analogue Potassium Acetate (LD 50 for male rats = 3.25 (2.48-4.26) g/kg) and the molecular weights, the read-across approach is applied and the LD50 for substance Ammonium Acetate is calculated to be 2.55 (1.95 -3.35) g/kg bw.

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: The analogue Potassium Acetate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the acute oral toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published data (no information on test method) on the analogue Potassium Acetate.

GLP compliance:

not specified

Test type:

other: read-across from study with an analogue

Sex:

not specified

Dose descriptor:

LD50

Effect level:

2 552.58 mg/kg bw

Based on:

test mat.

Other findings:

Based on the experimental results obtained with the analogue Potassium Acetate (LD 50 for rats = 3250 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for substance Ammonium Acetate is calculated to be 2552.58 mg/kg bw.

The analogue Potassium Acetate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties. These properties are:
- a low log Pow value which is -3.72 for Potassium Acetate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 2.5 g/mL at 20 ºC for Potassium Acetate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 98.14 for Potassium Acetate and 77.08 for Ammonium Acetate.

The LD50 for substance Ammonium Acetate is calculated to be 2552.58 mg/kg bw.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for substance Ammonium Acetate is calculated to be 2552.58 mg/kg bw.

Executive summary:

Based on the experimental results obtained (reported under the endpoint record 07.02.01_02 KAc) with the analogue Potassium Acetate (LD 50 for rats = 3250 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for substance Ammonium Acetate is calculated to be 2552.58 mg/kg bw.

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test method was similar to OECD guideline 423. No data on GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

not specified

Remarks:

(No data on control)

GLP compliance:

not specified

Test type:

acute toxic class method

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5-6 weeks old
- Fasting period before study: 16 hours
- Acclimation period: At least, 5 days.

Route of administration:

oral: unspecified

Vehicle:

water

Doses:

20, 200, and 2000 mg/kg bw

No. of animals per sex per dose:

Three animals of each sex per dose.

Control animals:

not specified

Details on study design:

Careful observations of animals, including body weight changes, mortality figures, gross lesion and behavioural and clinical abnormality, were performed for 14 days.
Necropsy of all animals was carried out at the end of the test.
Animal experiments were performed simultaneously in two different laboratories.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Clinical signs:

other: No clinical symptoms are reported.

The LD50 for Ammonium sulfate was equal or greater than 2000 mg/kg bw in male and female rats.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium sulfate was equal or greater than 2000 mg/kg bw in male and female rats.

Executive summary:

In a study similar to OECD guideline 423, a LD50 value of about 2000 mg/kg bw is reported for male and female rats.

Reference 6

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test method was similar to OECD guideline 423. No data on GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

not specified

Remarks:

(No data on control)

GLP compliance:

not specified

Test type:

acute toxic class method

Species:

mouse

Strain:

other: ddy

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5-6 weeks old
- Fasting period before study: 16 hours
- Acclimation period: At least, 5 days.

Route of administration:

oral: unspecified

Vehicle:

water

Doses:

20, 200, and 2000 mg/kg bw

No. of animals per sex per dose:

Three animals of each sex per dose.

Control animals:

not specified

Details on study design:

Careful observations of animals, including body weight changes, mortality figures, gross lesion and behavioural and clinical abnormality, were performed for 14 days.
Necropsy of all animals was carried out at the end of the test.
Animal experiments were performed simultaneously in two different laboratories.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Clinical signs:

other: No clinical symptoms are reported.

The LD50 for Ammonium sulfate was greater than 2000 mg/kg bw in male and female mice.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium sulfate was greater than 2000 mg/kg bw in male and female mice.

Executive summary:

In a study similar to OECD guideline 423, a LD50 value greater than 2000 mg/kg bw is reported for male and female mice.

Reference 7

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test method was similar to OECD guideline 401. No data on GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

not specified

Remarks:

(No data on control)

GLP compliance:

not specified

Test type:

standard acute method

Species:

mouse

Strain:

other: ddy

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5-6 weeks old
- Fasting period before study: 16 hours
- Acclimation period: At least, 5 days.

Route of administration:

oral: unspecified

Vehicle:

water

Doses:

20, 200, and 2000 mg/kg bw

No. of animals per sex per dose:

Three animals of each sex per dose.

Control animals:

not specified

Details on study design:

Careful observations of animals, including body weight changes, mortality figures, gross lesion and behavioural and clinical abnormality, were performed for 14 days.
Necropsy of all animals was carried out at the end of the test.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

3 040 mg/kg bw

Based on:

test mat.

95% CL:

2 670 - 3 440

Clinical signs:

other: No clinical symptoms are reported.

The LD50 for Ammonium sulfate was 3040 (2670 -3440) mg/kg bw in male and female mice.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium sulfate was 3040 (2670 -3440) mg/kg bw in male and female mice.

Executive summary:

In a full LD50 test according to the Toxicity Guidelines of Japan (1984) an oral LD50 for mice of 3040 (2670 -3440) mg/kg bw was obtained. Details on clinical signs and necropsy findings were not given.

Reference 8

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Ammonium Sulfate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the acute oral toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (test method similar to OECD 423) on the analogue Ammonium sulfate.

GLP compliance:

not specified

Test type:

other: read-across from an acute toxic class method with an analogue

Sex:

male/female

Dose descriptor:

LD50

Effect level:

>= 2 333.28 mg/kg bw

Based on:

test mat.

Other findings:

Based on the experimental results obtained with the analogue Ammonium sulfate (LD 50 for rats >= 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be >= 2333.28 mg/kg bw.

The analogue Ammonium Sulfate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the aquatic toxicity endpoint. These properties are:
- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

The LD50 for Ammonium Acetate is calculated to be >= 2333.28 mg/kg bw.

The analogue Ammonium Sulphate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties:

- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,

- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and

- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

As indicated in the European Chemical Agency Practical Guide 6 “How to report read –across and categories”, the structural grouping was realized using “OECD QSAR APPLICATION TOOL BOX” version 1.1.0. Presented results show that both substances have common (eco)toxicological behavior (attachment).

Table 1. Data Matrix, Analogue Approach.

CAS Number		Source chemical 7783-20-2	Target chemical 631-61-8
CHEMICAL NAME		Ammonium sulphate	Ammonium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Experimental data: Decomposes above 280 ºC	Experimental data: 114 ºC
Boiling Point		Experimental data: Decomposes above 280 ºC	Estimated data: 312.76 ºC
Density		Experimental data: 1.769 at 50 ºC	Experimental results: 1.07-1.17 g/cm3 at 20 ºC
Vapour Pressure		Experimental data: 4.053E-07 Pa (partial pressure of ammonia over solid (NH4)2SO4 at 25 ºC)	Estimated data: 0.02 Pa at 25 ºC
Partition Coefficient (log Kow)		Experimental data: –5.1 at 25 ºC	Estimated data: -2.79
Water solubility		Experimental data: 764 g/L at 20 ºC	Experimental data: 1480 g/L at 4 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental data: In unsterilized soil, ammonium sulfate is mineralized fairly rapidly, and subsequently nitrified. Nitrification and de-nitrification processes also occur naturally in streams and rivers, as well as in many secondary sewage treatment processes	Experimental results on Ammonium Acetate, read-across from experimental data on Sodium Acetate and read-across from estimated data on Ammonia and Acetic Acid, based on functional group:   Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		Experimental data: (96 h) LC50 > 100 mg/L (Pimephales promelas)	Experimental data and read-across from Potassium Acetate, based on molecular weights:   LC50 = 392.70 mg/L.
Acute Toxicity to Aquatic Invertebrates		Experimental data: (96 h) LC50 > 100 mg/L (Asellus Intermedius, Daphnia magna, Dugesia tigrina and Gammarus fasciatus)	Read-across from experimental data on analogues Sodium Acetate, Potassium Acetate and Ammonia, based on molecular weights:   EC50 = 108.81 - 939.66 mg/L
Toxicity to Aquatic Plants		Experimental data: (18 d) EC50 = ca. 25476 mg/L (ca. 2700 mg NH3/L) (Chlorella vulgaris)	Read-across from experimental data on analogues Acetic Acid, Potassium Acetate and Ammonium Sulphate, based on molecular weights: (72 h) EC50 > 392.70 mg/L; (72 h) NOEC = 392.70 mg/L.
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LC50 >= 2000 mg/kg bw (rat, mouse) LC50 = 3040 mg/kg bw (mouse)	Weight of evidence: Read-across from experimental data on Potassium Acetate and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-3546.59 mg/kg bw
Acute Toxicity: Inhalation		Experimental data: (4 h) LC50 > 3.6 mg/m3 (rats) (1 h) LC50 > 2 mg/m3 (rabbits)	No data
Acute Toxicity: Dermal		Experimental data:   LC50 > 2000 mg/kg bw (rats, mice)	Weight of evidence: Read-across from experimental data on Fumaric Acid and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-26556.42 mg/kg bw
Skin Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits. -         Single exposure for 20 hours, intact skin: no skin effects observed. -         Multiple exposures, 8 hrs/day for 5 consecutive days, intact skin: no skin effects observed.	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate and Ammonium Lactate, and Ammonium Stearate based on functional group: The substance Ammonium Acetate is considered as not irritating for skin.
Eye Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits (50 mm3 of undiluted substance).	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate, Ammonium Sulphate, and Ammonium Stearate, based on functional group: The substance Ammonium Acetate is considered as not irritating for eyes.
Skin Sensitization		No data	Weight of evidence:   Read-across approach from experimental results on Citric Acid, Glycolic Acid, Sodium Glycolate, Lactic Acid, Ammonium Lactate, and Triacetin, based on functional group:   All this substances were not sensitising for human and guinea pigs. Based on these results, Ammonium acetate is considered to be not sensitizing.
Repeated Dose Toxicity		Experimental data: A 14-day inhalation study on rats exposed to 300 mg/m3, the only tested dose, did not report histopathological changes in the lower respiratory tract. As the respiratory tract is the target organ for inhalation exposure, the NOEL for toxicity to the lower respiratory tract is 300 mg/m3.   The NOAEL after feeding diets containing ammonium sulfate for 13 weeks to rats was 886 mg/kg bw/day. The only toxicity sign found was diarrhea in male animals of the high-dose group (LOAEL: 1792 mg/kg bw/day).	Repeated dose toxicity: oral: Weight of evidence: Experimental results:   Repeated dose toxicity: oral: 90 days withfemale Wistar rats. The NOAEL was 3150.4 mg/kg bw/day. Repeated dose toxicity: oral: 15 days study with female Wistar rats. The NOAEL 3102.2 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights:   The NOAEL >= 0.047 mg/kg bw/day, in male rats chronically treated for 8 months via drinking water. The NOAEL >= 3382.76 mg/kg bw/day, in male Wistar rats daily treated for 4 weeks by feed. The NOAEL >= 19.73 mg/kg bw/day, in male Long-Evans rats treated for 3 months in the diet. The NOAEL >= 0.0094 mg/kg bw/day, in male Wistar rats treated by drinking water for 112 days.    Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   The NOAEL >= 54 mg/kg bw/day, in albino rats treated for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	Experimental data: Ammonium sulfate was not mutagenic in bacteria (Ames test) and yeasts with and without metabolic activation systems.	Weight of evidence:   Read-across from Sodium Acetate (category analogue) based on functional group:   Reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with metabolic activation. Resultslead to the conclusion that Ammonium Acetate did not cause point mutations in the microbial systems. Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered to be not mutagenic on S.typhimurium TA 98, TA 100, TA 1535, TA 97, and/or TA 1537, with and without metabolic activation. Read-across from experimental data on Ammonia, anhydrous, based on functional group: Ammonium acetate is considered to be not mutagenic on Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA, with and without metabolic activation.   Read-across from experimental data on Ammonia, aqueous solution, based on functional group: Ammonium acetate is considered not mutagenic on E. coli Sd-4-73, without metabolic activation.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Ammonium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation. Read-across from the analogue Phenoxy acetic acid, based on functional group: Ammonium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation.   Estimated data from Danish (Q)SAR Database: Ammonium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	-         Chromosomal aberration	Experimental data:   Ammonium sulfate did not induce chromosomal aberrations in mammalian or human cell cultures.	Weight of evidence: Read-across from Sodium Acetate (category analogue) based on functional group:   In an in vitro chromosomal aberration assay with a Chinese hamster fibroblast cell line, CHL, without metabolic activation systems, it is concluded that Ammonium acetate did not induce chromosomal aberrations(including gaps). Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation. Read-across from Ammonium Sulfate, based on functional group: Ammonium Acetate is not considered mutagenic on Chinese Hamster Ovary cells, in the absence of a metabolic activation system.
Genetic Toxicity in vivo		No data	Key studies: Read-across from Sodium Acetate (category analogue) based on functional group:   The Testicular DNA-synthesis inhibition test (DSI test) on male mice provides evidence that Ammonium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Test substance did not inhibit DNA replication in this assay.
Carcinogenicity		Experimental data: Similarly to other salts, high doses of ammonium sulfate may have the capability of tumor promotion in the rat stomach; it is, however, much less potent than sodium chloride when tested under identical conditions.	No data
Reproductive Toxicity		TOXICITY TO REPRODUCTION: Fischer 344 rats (10 rats/sex/dose) were exposed during 13 weeks to diets containing 0, 0.38, 0.75, 1.5 or 3 % ammonium sulfate (corresponding to 0, 222, 441, 886, 1792 mg/kg bw/day in males and to 0, 239, 484, 961, 1975 mg/kg bw/day in females). No effects on the reproductive organs were observed. Therefore, the NOAEL was 886 mg/kg bw/day for males and 1975 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: No data.	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). A fertility test on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 722.25 mg/kg bw/day, and LOAEL greater than 722.25 mg/kg bw/day for reproductive effects. Read-across from the analogue Citric Acid, sodium salt, based on molecular weights: A fertility study on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 54.0 mg/kg bw/day, and LOAEL greater than 54.0 mg/kg bw/day for reproductive effects. Read-across from the analogue Ammonium sulfate, based on molecular weights: A study on male and female rats exposed for 13 weeks to diets with Ammonium Sulfate. For Ammonium Acetate, the NOAEL is calculated to be 1033.64 mg/kg bw/day for males, and 2304.12 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Experimental results: A study on female rats fed an ammonium-containing diet starting on day 1 of pregnancy until weaning (at posnatal day on 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. The NOAEL for developmental toxicity was 4293 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights: Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Ammonium Acetate, theNOAEL is calculated to be939.66 mg/kg bw/day (based on maternal toxicity: mortality, pregnancy and resorption; and on neonatal effects: mortality and body weight). Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). Read-across from the analogue substance Calcium Formate, based on molecular weights: A three-generation drinking water study was performed. For Ammonium Acetate, the NOAEL is calculated to be equal or higher than 236.96 mg/kg bw/day. Read-across from Acetic Acid, based on molecular weights: A one-generation study was performed on female mice, rats and rabbits with Acetic Acid. The read-across approach was applied and the NOAEL with the substance Ammonium acetate is calculated to be equal or greater than 2055.47 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium Acetate is calculated to be >= 2333.28 mg/kg bw.

Executive summary:

Based on the experimental results (reported under the endpoint record 07.02.01_05 Ammonium sulfate) obtained with the analogue Ammonium sulfate (LD 50 for rats >= 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be >= 2333.28 mg/kg bw.

Reference 9

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Ammonium Sulfate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the acute oral toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (test method similar to OECD 423) on the analogue Ammonium sulfate.

GLP compliance:

not specified

Test type:

other: read-across from an acute toxic class method with an analogue

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 333.28 mg/kg bw

Based on:

test mat.

Other findings:

Based on the experimental results obtained with the analogue Ammonium sulfate (LD 50 for mice > 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

The analogue Ammonium Sulfate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the aquatic toxicity endpoint. These properties are:
- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

The LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

The analogue Ammonium Sulphate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties:

- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,

- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and

- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

As indicated in the European Chemical Agency Practical Guide 6 “How to report read –across and categories”, the structural grouping was realized using “OECD QSAR APPLICATION TOOL BOX” version 1.1.0. Presented results show that both substances have common (eco)toxicological behavior (attachment).

Table 1. Data Matrix, Analogue Approach.

CAS Number		Source chemical 7783-20-2	Target chemical 631-61-8
CHEMICAL NAME		Ammonium sulphate	Ammonium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Experimental data: Decomposes above 280 ºC	Experimental data: 114 ºC
Boiling Point		Experimental data: Decomposes above 280 ºC	Estimated data: 312.76 ºC
Density		Experimental data: 1.769 at 50 ºC	Experimental results: 1.07-1.17 g/cm3 at 20 ºC
Vapour Pressure		Experimental data: 4.053E-07 Pa (partial pressure of ammonia over solid (NH4)2SO4 at 25 ºC)	Estimated data: 0.02 Pa at 25 ºC
Partition Coefficient (log Kow)		Experimental data: –5.1 at 25 ºC	Estimated data: -2.79
Water solubility		Experimental data: 764 g/L at 20 ºC	Experimental data: 1480 g/L at 4 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental data: In unsterilized soil, ammonium sulfate is mineralized fairly rapidly, and subsequently nitrified. Nitrification and de-nitrification processes also occur naturally in streams and rivers, as well as in many secondary sewage treatment processes	Experimental results on Ammonium Acetate, read-across from experimental data on Sodium Acetate and read-across from estimated data on Ammonia and Acetic Acid, based on functional group:   Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		Experimental data: (96 h) LC50 > 100 mg/L (Pimephales promelas)	Experimental data and read-across from Potassium Acetate, based on molecular weights:   LC50 = 392.70 mg/L.
Acute Toxicity to Aquatic Invertebrates		Experimental data: (96 h) LC50 > 100 mg/L (Asellus Intermedius, Daphnia magna, Dugesia tigrina and Gammarus fasciatus)	Read-across from experimental data on analogues Sodium Acetate, Potassium Acetate and Ammonia, based on molecular weights:   EC50 = 108.81 - 939.66 mg/L
Toxicity to Aquatic Plants		Experimental data: (18 d) EC50 = ca. 25476 mg/L (ca. 2700 mg NH3/L) (Chlorella vulgaris)	Read-across from experimental data on analogues Acetic Acid, Potassium Acetate and Ammonium Sulphate, based on molecular weights: (72 h) EC50 > 392.70 mg/L; (72 h) NOEC = 392.70 mg/L.
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LC50 >= 2000 mg/kg bw (rat, mouse) LC50 = 3040 mg/kg bw (mouse)	Weight of evidence: Read-across from experimental data on Potassium Acetate and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-3546.59 mg/kg bw
Acute Toxicity: Inhalation		Experimental data: (4 h) LC50 > 3.6 mg/m3 (rats) (1 h) LC50 > 2 mg/m3 (rabbits)	No data
Acute Toxicity: Dermal		Experimental data:   LC50 > 2000 mg/kg bw (rats, mice)	Weight of evidence: Read-across from experimental data on Fumaric Acid and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-26556.42 mg/kg bw
Skin Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits. -         Single exposure for 20 hours, intact skin: no skin effects observed. -         Multiple exposures, 8 hrs/day for 5 consecutive days, intact skin: no skin effects observed.	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate and Ammonium Lactate, and Ammonium Stearate based on functional group: The substance Ammonium Acetate is considered as not irritating for skin.
Eye Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits (50 mm3 of undiluted substance).	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate, Ammonium Sulphate, and Ammonium Stearate, based on functional group: The substance Ammonium Acetate is considered as not irritating for eyes.
Skin Sensitization		No data	Weight of evidence:   Read-across approach from experimental results on Citric Acid, Glycolic Acid, Sodium Glycolate, Lactic Acid, Ammonium Lactate, and Triacetin, based on functional group:   All this substances were not sensitising for human and guinea pigs. Based on these results, Ammonium acetate is considered to be not sensitizing.
Repeated Dose Toxicity		Experimental data: A 14-day inhalation study on rats exposed to 300 mg/m3, the only tested dose, did not report histopathological changes in the lower respiratory tract. As the respiratory tract is the target organ for inhalation exposure, the NOEL for toxicity to the lower respiratory tract is 300 mg/m3.   The NOAEL after feeding diets containing ammonium sulfate for 13 weeks to rats was 886 mg/kg bw/day. The only toxicity sign found was diarrhea in male animals of the high-dose group (LOAEL: 1792 mg/kg bw/day).	Repeated dose toxicity: oral: Weight of evidence: Experimental results:   Repeated dose toxicity: oral: 90 days withfemale Wistar rats. The NOAEL was 3150.4 mg/kg bw/day. Repeated dose toxicity: oral: 15 days study with female Wistar rats. The NOAEL 3102.2 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights:   The NOAEL >= 0.047 mg/kg bw/day, in male rats chronically treated for 8 months via drinking water. The NOAEL >= 3382.76 mg/kg bw/day, in male Wistar rats daily treated for 4 weeks by feed. The NOAEL >= 19.73 mg/kg bw/day, in male Long-Evans rats treated for 3 months in the diet. The NOAEL >= 0.0094 mg/kg bw/day, in male Wistar rats treated by drinking water for 112 days.    Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   The NOAEL >= 54 mg/kg bw/day, in albino rats treated for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	Experimental data: Ammonium sulfate was not mutagenic in bacteria (Ames test) and yeasts with and without metabolic activation systems.	Weight of evidence:   Read-across from Sodium Acetate (category analogue) based on functional group:   Reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with metabolic activation. Resultslead to the conclusion that Ammonium Acetate did not cause point mutations in the microbial systems. Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered to be not mutagenic on S.typhimurium TA 98, TA 100, TA 1535, TA 97, and/or TA 1537, with and without metabolic activation. Read-across from experimental data on Ammonia, anhydrous, based on functional group: Ammonium acetate is considered to be not mutagenic on Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA, with and without metabolic activation.   Read-across from experimental data on Ammonia, aqueous solution, based on functional group: Ammonium acetate is considered not mutagenic on E. coli Sd-4-73, without metabolic activation.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Ammonium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation. Read-across from the analogue Phenoxy acetic acid, based on functional group: Ammonium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation.   Estimated data from Danish (Q)SAR Database: Ammonium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	-         Chromosomal aberration	Experimental data:   Ammonium sulfate did not induce chromosomal aberrations in mammalian or human cell cultures.	Weight of evidence: Read-across from Sodium Acetate (category analogue) based on functional group:   In an in vitro chromosomal aberration assay with a Chinese hamster fibroblast cell line, CHL, without metabolic activation systems, it is concluded that Ammonium acetate did not induce chromosomal aberrations(including gaps). Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation. Read-across from Ammonium Sulfate, based on functional group: Ammonium Acetate is not considered mutagenic on Chinese Hamster Ovary cells, in the absence of a metabolic activation system.
Genetic Toxicity in vivo		No data	Key studies: Read-across from Sodium Acetate (category analogue) based on functional group:   The Testicular DNA-synthesis inhibition test (DSI test) on male mice provides evidence that Ammonium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Test substance did not inhibit DNA replication in this assay.
Carcinogenicity		Experimental data: Similarly to other salts, high doses of ammonium sulfate may have the capability of tumor promotion in the rat stomach; it is, however, much less potent than sodium chloride when tested under identical conditions.	No data
Reproductive Toxicity		TOXICITY TO REPRODUCTION: Fischer 344 rats (10 rats/sex/dose) were exposed during 13 weeks to diets containing 0, 0.38, 0.75, 1.5 or 3 % ammonium sulfate (corresponding to 0, 222, 441, 886, 1792 mg/kg bw/day in males and to 0, 239, 484, 961, 1975 mg/kg bw/day in females). No effects on the reproductive organs were observed. Therefore, the NOAEL was 886 mg/kg bw/day for males and 1975 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: No data.	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). A fertility test on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 722.25 mg/kg bw/day, and LOAEL greater than 722.25 mg/kg bw/day for reproductive effects. Read-across from the analogue Citric Acid, sodium salt, based on molecular weights: A fertility study on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 54.0 mg/kg bw/day, and LOAEL greater than 54.0 mg/kg bw/day for reproductive effects. Read-across from the analogue Ammonium sulfate, based on molecular weights: A study on male and female rats exposed for 13 weeks to diets with Ammonium Sulfate. For Ammonium Acetate, the NOAEL is calculated to be 1033.64 mg/kg bw/day for males, and 2304.12 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Experimental results: A study on female rats fed an ammonium-containing diet starting on day 1 of pregnancy until weaning (at posnatal day on 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. The NOAEL for developmental toxicity was 4293 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights: Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Ammonium Acetate, theNOAEL is calculated to be939.66 mg/kg bw/day (based on maternal toxicity: mortality, pregnancy and resorption; and on neonatal effects: mortality and body weight). Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). Read-across from the analogue substance Calcium Formate, based on molecular weights: A three-generation drinking water study was performed. For Ammonium Acetate, the NOAEL is calculated to be equal or higher than 236.96 mg/kg bw/day. Read-across from Acetic Acid, based on molecular weights: A one-generation study was performed on female mice, rats and rabbits with Acetic Acid. The read-across approach was applied and the NOAEL with the substance Ammonium acetate is calculated to be equal or greater than 2055.47 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

Executive summary:

Based on the experimental results (reported under the endpoint record 07.02.01_06 Ammonium sulfate) obtained with the analogue Ammonium sulfate (LD 50 for mice > 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

Reference 10

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Ammonium Sulfate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the acute oral toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (test method similar to OECD 401) on the analogue Ammonium sulfate.

GLP compliance:

not specified

Test type:

other: read-across from a standard acute method with an analogue

Sex:

male/female

Dose descriptor:

LD50

Effect level:

3 546.59 mg/kg bw

Based on:

test mat.

95% CL:

3 114.93 - 4 013.25

Other findings:

Based on the experimental results obtained with the analogue Ammonium sulfate (LD 50 for mice = 3040 (2670-3440) mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be 3546.59 (3114.93-4013.25) mg/kg bw.

The analogue Ammonium Sulfate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the aquatic toxicity endpoint. These properties are:
- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

The LD50 for Ammonium Acetate is calculated to be 3546.59 (3114.93-4013.25) mg/kg bw.

The analogue Ammonium Sulphate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties:

- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,

- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and

- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

As indicated in the European Chemical Agency Practical Guide 6 “How to report read –across and categories”, the structural grouping was realized using “OECD QSAR APPLICATION TOOL BOX” version 1.1.0. Presented results show that both substances have common (eco)toxicological behavior (attachment).

Table 1. Data Matrix, Analogue Approach.

CAS Number		Source chemical 7783-20-2	Target chemical 631-61-8
CHEMICAL NAME		Ammonium sulphate	Ammonium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Experimental data: Decomposes above 280 ºC	Experimental data: 114 ºC
Boiling Point		Experimental data: Decomposes above 280 ºC	Estimated data: 312.76 ºC
Density		Experimental data: 1.769 at 50 ºC	Experimental results: 1.07-1.17 g/cm3 at 20 ºC
Vapour Pressure		Experimental data: 4.053E-07 Pa (partial pressure of ammonia over solid (NH4)2SO4 at 25 ºC)	Estimated data: 0.02 Pa at 25 ºC
Partition Coefficient (log Kow)		Experimental data: –5.1 at 25 ºC	Estimated data: -2.79
Water solubility		Experimental data: 764 g/L at 20 ºC	Experimental data: 1480 g/L at 4 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental data: In unsterilized soil, ammonium sulfate is mineralized fairly rapidly, and subsequently nitrified. Nitrification and de-nitrification processes also occur naturally in streams and rivers, as well as in many secondary sewage treatment processes	Experimental results on Ammonium Acetate, read-across from experimental data on Sodium Acetate and read-across from estimated data on Ammonia and Acetic Acid, based on functional group:   Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		Experimental data: (96 h) LC50 > 100 mg/L (Pimephales promelas)	Experimental data and read-across from Potassium Acetate, based on molecular weights:   LC50 = 392.70 mg/L.
Acute Toxicity to Aquatic Invertebrates		Experimental data: (96 h) LC50 > 100 mg/L (Asellus Intermedius, Daphnia magna, Dugesia tigrina and Gammarus fasciatus)	Read-across from experimental data on analogues Sodium Acetate, Potassium Acetate and Ammonia, based on molecular weights:   EC50 = 108.81 - 939.66 mg/L
Toxicity to Aquatic Plants		Experimental data: (18 d) EC50 = ca. 25476 mg/L (ca. 2700 mg NH3/L) (Chlorella vulgaris)	Read-across from experimental data on analogues Acetic Acid, Potassium Acetate and Ammonium Sulphate, based on molecular weights: (72 h) EC50 > 392.70 mg/L; (72 h) NOEC = 392.70 mg/L.
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LC50 >= 2000 mg/kg bw (rat, mouse) LC50 = 3040 mg/kg bw (mouse)	Weight of evidence: Read-across from experimental data on Potassium Acetate and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-3546.59 mg/kg bw
Acute Toxicity: Inhalation		Experimental data: (4 h) LC50 > 3.6 mg/m3 (rats) (1 h) LC50 > 2 mg/m3 (rabbits)	No data
Acute Toxicity: Dermal		Experimental data:   LC50 > 2000 mg/kg bw (rats, mice)	Weight of evidence: Read-across from experimental data on Fumaric Acid and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-26556.42 mg/kg bw
Skin Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits. -         Single exposure for 20 hours, intact skin: no skin effects observed. -         Multiple exposures, 8 hrs/day for 5 consecutive days, intact skin: no skin effects observed.	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate and Ammonium Lactate, and Ammonium Stearate based on functional group: The substance Ammonium Acetate is considered as not irritating for skin.
Eye Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits (50 mm3 of undiluted substance).	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate, Ammonium Sulphate, and Ammonium Stearate, based on functional group: The substance Ammonium Acetate is considered as not irritating for eyes.
Skin Sensitization		No data	Weight of evidence:   Read-across approach from experimental results on Citric Acid, Glycolic Acid, Sodium Glycolate, Lactic Acid, Ammonium Lactate, and Triacetin, based on functional group:   All this substances were not sensitising for human and guinea pigs. Based on these results, Ammonium acetate is considered to be not sensitizing.
Repeated Dose Toxicity		Experimental data: A 14-day inhalation study on rats exposed to 300 mg/m3, the only tested dose, did not report histopathological changes in the lower respiratory tract. As the respiratory tract is the target organ for inhalation exposure, the NOEL for toxicity to the lower respiratory tract is 300 mg/m3.   The NOAEL after feeding diets containing ammonium sulfate for 13 weeks to rats was 886 mg/kg bw/day. The only toxicity sign found was diarrhea in male animals of the high-dose group (LOAEL: 1792 mg/kg bw/day).	Repeated dose toxicity: oral: Weight of evidence: Experimental results:   Repeated dose toxicity: oral: 90 days withfemale Wistar rats. The NOAEL was 3150.4 mg/kg bw/day. Repeated dose toxicity: oral: 15 days study with female Wistar rats. The NOAEL 3102.2 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights:   The NOAEL >= 0.047 mg/kg bw/day, in male rats chronically treated for 8 months via drinking water. The NOAEL >= 3382.76 mg/kg bw/day, in male Wistar rats daily treated for 4 weeks by feed. The NOAEL >= 19.73 mg/kg bw/day, in male Long-Evans rats treated for 3 months in the diet. The NOAEL >= 0.0094 mg/kg bw/day, in male Wistar rats treated by drinking water for 112 days.    Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   The NOAEL >= 54 mg/kg bw/day, in albino rats treated for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	Experimental data: Ammonium sulfate was not mutagenic in bacteria (Ames test) and yeasts with and without metabolic activation systems.	Weight of evidence:   Read-across from Sodium Acetate (category analogue) based on functional group:   Reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with metabolic activation. Resultslead to the conclusion that Ammonium Acetate did not cause point mutations in the microbial systems. Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered to be not mutagenic on S.typhimurium TA 98, TA 100, TA 1535, TA 97, and/or TA 1537, with and without metabolic activation. Read-across from experimental data on Ammonia, anhydrous, based on functional group: Ammonium acetate is considered to be not mutagenic on Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA, with and without metabolic activation.   Read-across from experimental data on Ammonia, aqueous solution, based on functional group: Ammonium acetate is considered not mutagenic on E. coli Sd-4-73, without metabolic activation.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Ammonium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation. Read-across from the analogue Phenoxy acetic acid, based on functional group: Ammonium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation.   Estimated data from Danish (Q)SAR Database: Ammonium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	-         Chromosomal aberration	Experimental data:   Ammonium sulfate did not induce chromosomal aberrations in mammalian or human cell cultures.	Weight of evidence: Read-across from Sodium Acetate (category analogue) based on functional group:   In an in vitro chromosomal aberration assay with a Chinese hamster fibroblast cell line, CHL, without metabolic activation systems, it is concluded that Ammonium acetate did not induce chromosomal aberrations(including gaps). Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation. Read-across from Ammonium Sulfate, based on functional group: Ammonium Acetate is not considered mutagenic on Chinese Hamster Ovary cells, in the absence of a metabolic activation system.
Genetic Toxicity in vivo		No data	Key studies: Read-across from Sodium Acetate (category analogue) based on functional group:   The Testicular DNA-synthesis inhibition test (DSI test) on male mice provides evidence that Ammonium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Test substance did not inhibit DNA replication in this assay.
Carcinogenicity		Experimental data: Similarly to other salts, high doses of ammonium sulfate may have the capability of tumor promotion in the rat stomach; it is, however, much less potent than sodium chloride when tested under identical conditions.	No data
Reproductive Toxicity		TOXICITY TO REPRODUCTION: Fischer 344 rats (10 rats/sex/dose) were exposed during 13 weeks to diets containing 0, 0.38, 0.75, 1.5 or 3 % ammonium sulfate (corresponding to 0, 222, 441, 886, 1792 mg/kg bw/day in males and to 0, 239, 484, 961, 1975 mg/kg bw/day in females). No effects on the reproductive organs were observed. Therefore, the NOAEL was 886 mg/kg bw/day for males and 1975 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: No data.	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). A fertility test on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 722.25 mg/kg bw/day, and LOAEL greater than 722.25 mg/kg bw/day for reproductive effects. Read-across from the analogue Citric Acid, sodium salt, based on molecular weights: A fertility study on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 54.0 mg/kg bw/day, and LOAEL greater than 54.0 mg/kg bw/day for reproductive effects. Read-across from the analogue Ammonium sulfate, based on molecular weights: A study on male and female rats exposed for 13 weeks to diets with Ammonium Sulfate. For Ammonium Acetate, the NOAEL is calculated to be 1033.64 mg/kg bw/day for males, and 2304.12 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Experimental results: A study on female rats fed an ammonium-containing diet starting on day 1 of pregnancy until weaning (at posnatal day on 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. The NOAEL for developmental toxicity was 4293 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights: Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Ammonium Acetate, theNOAEL is calculated to be939.66 mg/kg bw/day (based on maternal toxicity: mortality, pregnancy and resorption; and on neonatal effects: mortality and body weight). Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). Read-across from the analogue substance Calcium Formate, based on molecular weights: A three-generation drinking water study was performed. For Ammonium Acetate, the NOAEL is calculated to be equal or higher than 236.96 mg/kg bw/day. Read-across from Acetic Acid, based on molecular weights: A one-generation study was performed on female mice, rats and rabbits with Acetic Acid. The read-across approach was applied and the NOAEL with the substance Ammonium acetate is calculated to be equal or greater than 2055.47 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium Acetate is calculated to be 3546.59 (3114.93-4013.25) mg/kg bw.

Executive summary:

Based on the experimental results (reported under the endpoint record 07.02.01_07 Ammonium sulfate) obtained with the analogue Ammonium sulfate (LD 50 for mice = 3040 (2670-3440) mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be 3546.59 (3114.93-4013.25) mg/kg bw.

Endpoint conclusion

Dose descriptor:

LD50

Value:

2 338.28 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
According to REACH Annex VIII, column 2: In addition to the oral route (Annex VII, 8.5.1.), for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. The information is provided for dermal route.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Fumaric acid which shares the same functional group with Ammonium acetate, also has comparable values for the relevant molecular properties for acute dermal toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (study with a test method similar to OECD 402) on the analogue Fumaric Acid.

GLP compliance:

no

Test type:

other: read-across from a standard acute method with an analogue

Sex:

female

Dose descriptor:

LD50

Effect level:

> 26 556.42 mg/kg bw

Based on:

test mat.

Other findings:

Based on the experimental results obtained with the analogue Fumaric Acid (4h-LD 50 for New Zealand rabbits > 20000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD 50 for substance Ammonium Acetate is calculated to be greater than 26556.42 mg/kg bw under test conditions.

The analogue Fumaric Acid, which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties. These properties are:
- a low log Pow value which is 0.25 for Fumaric Acid and - 2.79 for Ammonium Acetate,
- water solubility which is 0.0063 g/mL at 25 ºC for Fumaric Acid and 1480 g/L at 4 ºC for Ammonium acetate, and
- molecular weights which are 116.07 for Fumaric Acid and 77.08 for Ammonium acetate.

The analogue Fumaric Acid which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties. These properties are:

- a low log Pow value which is 0.25 for Fumaric Acid and -2.79 for Ammonium Acetate,

- similar molecular weights which are 116.07 for Fumaric Acid and 77.08 for Ammonium Acetate.

Both chemicals are grouped together by US EPA category group Carboxylic Food Acids and Salts Category.

As indicated in the European Chemical Agency Practical Guide 6 “How to report read –across and categories”, the structural grouping was realized using “OECD QSAR APPLICATION TOOL BOX” version 1.1.0.Presented results show that both substances have common (eco)toxicological behavior (attachment).

Table 1. Data Matrix, Analogue Approach.

CAS Number		Source chemical 110-17-8	Target chemical 631-61-8
CHEMICAL NAME		Fumaric acid	Ammonium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Experimental results: 290°C (sublimes)	Experimental data: 114 ºC
Boiling Point		Experimental results: 290 °C (sublimes)	Estimated data: 312.76 ºC
Density		Experimental results: 1.64 at 20 ºC	Experimental results: 1.07-1.17 g/cm3 at 20 ºC
Vapour Pressure		Measured data: 1.5×10-4 mm Hg at 25°C	Estimated data: 0.02 Pa at 25 ºC
Partition Coefficient (log Kow)		Estimated data: 0.25	Estimated data: -2.79
Water solubility		Measured data: 0.0063 g/mL at 25 ºC	Experimental results: 1480 g/L at 4 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental results: Readily biodegradable	Experimental results on Ammonium Acetate, read-across from experimental data on Sodium Acetate and read-across from estimated data on Ammonia and Acetic Acid, based on functional group:   Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		No data	Experimental data and read-across from Potassium Acetate, based on molecular weights:   LC50 = 392.70 mg/L.
Acute Toxicity to Aquatic Invertebrates		Experimental data: 48-h EC50 = 212 mg/L (Daphnia magna)	Read-across from experimental data on analogues Sodium Acetate, Potassium Acetate and Ammonia, based on molecular weights:   EC50 = 108.81 - 939.66 mg/L
Toxicity to Aquatic Plants		Experimental data:   72-h EC50 (growth) = 41 mg/L(Scenedesmus subcapitata)	Read-across from experimental data on analogues Acetic Acid, Potassium Acetate and Ammonium Sulphate, based on molecular weights: (72 h) EC50 > 392.70 mg/L; (72 h) NOEC = 392.70 mg/L.
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LD 50 = 9300 - 10700 mg/kg bw (rats)	Weight of evidence: Read-across from experimental data on Potassium Acetate and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-3546.59 mg/kg bw
Acute Toxicity: Inhalation		No data	No data
Acute Toxicity: Dermal		Key study: LD50 (4 h) > 20000 mg/kg bw (female New Zealand White rabbits)	Weight of evidence: Read-across from experimental data on Fumaric Acid and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-26556.42 mg/kg bw
Skin Irritation/Corrosion		No data	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate and Ammonium Lactate, and Ammonium Stearate based on functional group: The substance Ammonium Acetate is considered as not irritating for skin.
Eye Irritation/Corrosion		Experimental data:   Fumaric Acid has been tested by application of a drop of 10% solution to the eyes of rabbits after mechanical removal of corneal epithelium to facilitate penetration, but it appeared to do no damage, & healing was similar to that in control eyes without test chemical.	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate, Ammonium Sulphate, and Ammonium Stearate, based on functional group: The substance Ammonium Acetate is considered as not irritating for eyes.
Skin sensitisation		No data	Weight of evidence:   Read-across approach from experimental results on Citric Acid, Glycolic Acid, Sodium Glycolate, Lactic Acid, Ammonium Lactate, and Triacetin, based on functional group:   All this substances were not sensitising for human and guinea pigs. Based on these results, Ammonium acetate is considered to be not sensitizing.
Repeated Dose Toxicity		Repeated dose toxicity: oral: Experimental data: Repeated dose toxicity: oral: 2-year study in male and female rats which were treated by diet. The LOAEL = 750 mg/kg bw/day (based on slight increases in mortality and increased incidence of testes degeneration at the highest dose tested). The NOAEL = 600 mg/kg bw/day.	Repeated dose toxicity: oral: Weight of evidence: Experimental results:   Repeated dose toxicity: oral: 90 days withfemale Wistar rats. The NOAEL was 3150.4 mg/kg bw/day . Repeated dose toxicity: oral: 15 days study with female Wistar rats. The NOAEL 3102.2 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights:   The NOAEL >= 0.047 mg/kg bw/day, in male rats chronically treated for 8 months via drinking water. The NOAEL >= 3382.76 mg/kg bw/day, in male Wistar rats daily treated for 4 weeks by feed. The NOAEL >= 19.73 mg/kg bw/day, in male Long-Evans rats treated for 3 months in the diet. The NOAEL >= 0.0094 mg/kg bw/day, in male Wistar rats treated by drinking water for 112 days.    Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   The NOAEL >= 54 mg/kg bw/day, in albino rats treated for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	In a bacterial reverse mutation assay usingS. typhimurium(TA98, TA100, TA1535, TA97 and TA1537) in the absence of metabolic activation and concentrations up to 1000μg/plate, fumaric acid was not mutagenic.	Weight of evidence:   Read-across from Sodium Acetate (category analogue) based on functional group:   Reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with metabolic activation. Resultslead to the conclusion that Ammonium Acetate did not cause point mutations in the microbial systems. Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered to be not mutagenic on S.typhimurium TA 98, TA 100, TA 1535, TA 97, and/or TA 1537, with and without metabolic activation. Read-across from experimental data on Ammonia, anhydrous, based on functional group: Ammonium acetate is considered to be not mutagenic on Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA, with and without metabolic activation.   Read-across from experimental data on Ammonia, aqueous solution, based on functional group: Ammonium acetate is considered not mutagenic on E. coli Sd-4-73, without metabolic activation.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Ammonium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation. Read-across from the analogue Phenoxy acetic acid, based on functional group: Ammonium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation.   Estimated data from Danish (Q)SAR Database: Ammonium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	Chromosomal aberration	Fumaric acid was assayed in anin vitroassay using Chinese hamster fibroblast cells in the absence of metabolic activation at doses up to 1 mg/mL; however, insufficient information was provided in the robust summary to adequately evaluate this study.	Weight of evidence: Read-across from Sodium Acetate (category analogue) based on functional group:   In an in vitro chromosomal aberration assay with a Chinese hamster fibroblast cell line, CHL, without metabolic activation systems, it is concluded that Ammonium acetate did not induce chromosomal aberrations(including gaps). Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation. Read-across from Ammonium Sulfate, based on functional group: Ammonium Acetate is not considered mutagenic on Chinese Hamster Ovary cells, in the absence of a metabolic activation system.
Genetic Toxicity in vivo		No data	Key studies: Read-across from Sodium Acetate (category analogue) based on functional group:   The Testicular DNA-synthesis inhibition test (DSI test) on male mice provides evidence that Ammonium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Test substance did not inhibit DNA replication in this assay.
Carcinogenicity		No data	No data
Reproductive Toxicity		No data	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). A fertility test on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 722.25 mg/kg bw/day, and LOAEL greater than 722.25 mg/kg bw/day for reproductive effects. Read-across from the analogue Citric Acid, sodium salt, based on molecular weights: A fertility study on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 54.0 mg/kg bw/day, and LOAEL greater than 54.0 mg/kg bw/day for reproductive effects. Read-across from the analogue Ammonium sulfate, based on molecular weights: A study on male and female rats exposed for 13 weeks to diets with Ammonium Sulfate. For Ammonium Acetate, the NOAEL is calculated to be 1033.64 mg/kg bw/day for males, and 2304.12 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Experimental results: A study on female rats fed an ammonium-containing diet starting on day 1 of pregnancy until weaning (at posnatal day on 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. The NOAEL for developmental toxicity was 4293 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights: Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Ammonium Acetate, theNOAEL is calculated to be939.66 mg/kg bw/day (based on maternal toxicity: mortality, pregnancy and resorption; and on neonatal effects: mortality and body weight). Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). Read-across from the analogue substance Calcium Formate, based on molecular weights: A three-generation drinking water study was performed. For Ammonium Acetate, the NOAEL is calculated to be equal or higher than 236.96 mg/kg bw/day. Read-across from Acetic Acid, based on molecular weights: A one-generation study was performed on female mice, rats and rabbits with Acetic Acid. The read-across approach was applied and the NOAEL with the substance Ammonium acetate is calculated to be equal or greater than 2055.47 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The (4h) LD 50 for substance Ammonium Acetate is calculated to be greater than 26556.42 mg/kg bw for rabbits.

Executive summary:

Based on the experimental results (reported under the endpoint record 07.02.03_02 Fumaric Acid) obtained with the analogue Fumaric Acid (4h-LD 50 for New Zealand rabbits > 20000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD 50 for substance Ammonium acetate is calculated to be higher than 26556.42 mg/kg bw under test conditions.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test method was similar to OECD 402 guideline, but only three animals per dose were used. No GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

(Only 3 female rabbits were used per dose)

GLP compliance:

no

Test type:

standard acute method

Species:

rabbit

Strain:

New Zealand White

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 3-4 kg bw
- Diet (e.g. ad libitum): Purina Rabbit Chow

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Type of wrap if used: Test substance was kept in place by gauze patches under a latex rubber film.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL

Duration of exposure:

4 hours

Doses:

No data on doses (the highest one was 20000 mg/kg bw)

No. of animals per sex per dose:

3 female New Zealand rabbits were used per dose.

Control animals:

not specified

Sex:

female

Dose descriptor:

LD50

Effect level:

> 20 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed at the highest dose of 20000 mg/kg bw

The LD50 for Fumaric Acid is higher than 20000 mg/kg bw for rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Fumaric Acid is higher than 20000 mg/kg bw for rabbits.

Executive summary:

The acute dermal toxicity of Fumaric Acid has been studied on Rabbits were clipped of all possible hair on back and flanks 24 hours before exposure. A 0.5 mL dose of Fumaric Acid was administered via a single skin penetration to three female albino New Zealand rabbits and kept in place by gauze patches under a latex rubber film. The patches remained in place for 4 hours. During that time the rabbits were fitted with leather restraining collars to prevent disturbance of the patch test. After 4 hours, the wrap and patches were removed and the test area was observed for visible tissue destruction.

No mortality was observed at the high dose of 20000 mg/kg b.w, so the LD50 is higher than 20000 mg/kg bw.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test method according to the Toxicity Guidelines of Japan (Ministry of Health and Welfare in Japan, 1984). No data on GLP.

Qualifier:

according to guideline

Guideline:

other: Toxicity Guidelines of Japan (Ministry of Health and Welfare in Japan, 1984).

Deviations:

not specified

GLP compliance:

not specified

Test type:

fixed dose procedure

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5-6 weeks old
- Housing: All animals were individually housed in stainless-steel cages.
- Acclimation period: At least, 5 days.

Type of coverage:

open

Vehicle:

other: water-acetone solution

Details on dermal exposure:

In the dermal toxicity test, hair was first removed from an area of 3 x 4 cm2 on the back of animals with an electric hair clipper, and then the chemical substance was dissolved in acetone and water and applied in a single dose to the skin surface of the clipped backs of the animals. The application sites were not covered but the treated areas were prevented from being licked by using a plastic collar or by fixing the animals on a plastic plate, and all animals were individually housed in stainless-steel cages.

Duration of exposure:

A single administration, but no data on removing.

Doses:

20, 200, and 2000 mg/kg bw.

No. of animals per sex per dose:

Three animals of each sex per dose.

Control animals:

not specified

Details on study design:

Careful observations of animals, including body weight changes, mortality figures, gross lesion and behavioural and clinical abnormality, were performed for 14 days.
Necropsy of all animals was carried out at the end of the test.
Animal experiments were performed simultaneously in two different laboratories.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Clinical signs:

other: No treatment-related clinical signs and necropsy findings were observed.

The LD50 was higher than 2000 mg/kg bw for rats after dermal application of ammonium sulfate.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 was greater than 2000 mg/kg bw for rats after dermal application of ammonium sulfate.

Executive summary:

In the dermal toxicity test, hair was first removed from an area of 3 x 4 cm2 on the back of rats with an electric hair clipper, and then the chemical substance was dissolved in acetone and water and applied in a single dose to the skin surface of the clipped backs of the animals. The application sites were not covered but the treated areas were prevented from being licked by using a plastic collar or by fixing the animals on a plastic plate, and all animals were individually housed in stainless-steel cages.

The LD50 values higher than 2000 mg/kg bw are reported for rats after dermal application of ammonium sulfate. No treatment-related clinical signs and necropsy findings were observed.

Reference 4

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test method according to the Toxicity Guidelines of Japan (Ministry of Health and Welfare in Japan, 1984). No data on GLP.

Qualifier:

according to guideline

Guideline:

other: Toxicity Guidelines of Japan (Ministry of Health and Welfare in Japan, 1984).

Deviations:

not specified

GLP compliance:

not specified

Test type:

fixed dose procedure

Species:

mouse

Strain:

other: ddy

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5-6 weeks old
- Housing: All animals were individually housed in stainless-steel cages.
- Acclimation period: At least, 5 days.

Type of coverage:

open

Vehicle:

other: water-acetone solution

Details on dermal exposure:

In the dermal toxicity test, hair was first removed from an area of 1 x 2 cm2 on the back of animals with an electric hair clipper, and then the chemical substance was dissolved in acetone and water and applied in a single dose to the skin surface of the clipped backs of the animals. The application sites were not covered but the treated areas were prevented from being licked by using a plastic collar or by fixing the animals on a plastic plate, and all animals were individually housed in stainless-steel cages.

Duration of exposure:

A single administration, but no data on removing.

Doses:

20, 200, and 2000 mg/kg bw.

No. of animals per sex per dose:

Three animals of each sex per dose.

Control animals:

not specified

Details on study design:

Careful observations of animals, including body weight changes, mortality figures, gross lesion and behavioural and clinical abnormality, were performed for 14 days.
Necropsy of all animals was carried out at the end of the test.
Animal experiments were performed simultaneously in two different laboratories.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Clinical signs:

other: No treatment-related clinical signs and necropsy findings were observed.

The LD50 was greater than 2000 mg/kg bw for mice after dermal application of ammonium sulfate.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 was greater than 2000 mg/kg bw for mice after dermal application of ammonium sulfate.

Executive summary:

In the dermal toxicity test, hair was first removed from an area of 1 x 2 cm2 on the back of mice with an electric hair clipper, and then the chemical substance was dissolved in acetone and water and applied in a single dose to the skin surface of the clipped backs of the animals. The application sites were not covered but the treated areas were prevented from being licked by using a plastic collar or by fixing the animals on a plastic plate, and all animals were individually housed in stainless-steel cages.

The LD50 values higher than 2000 mg/kg bw are reported for mice after dermal application of ammonium sulfate. No treatment-related clinical signs and necropsy findings were observed.

Reference 5

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Ammonium Sulfate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for acute dermal toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (study with a test method according to the Toxicity Guidelines of Japan (Ministry of Health and Welfare in Japan, 1984)) on the analogue Ammoniun sulphate.

GLP compliance:

not specified

Test type:

other: read-across from a fixed dose procedure with an analogue

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 333.28 mg/kg bw

Based on:

test mat.

Other findings:

Based on the experimental results obtained with the analogue Ammonium sulfate (LD 50 for rats > 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

The analogue Ammonium Sulfate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the aquatic toxicity endpoint. These properties are:
- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

The analogue Ammonium Sulphate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties:

- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,

- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and

- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

As indicated in the European Chemical Agency Practical Guide 6 “How to report read –across and categories”, the structural grouping was realized using “OECD QSAR APPLICATION TOOL BOX” version 1.1.0. Presented results show that both substances have common (eco)toxicological behavior (attachment).

Table 1. Data Matrix, Analogue approach.

CAS Number		Source chemical 7783-20-2	Target chemical 631-61-8
CHEMICAL NAME		Ammonium sulphate	Ammonium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Experimental data: Decomposes above 280 ºC	Experimental data: 114 ºC
Boiling Point		Experimental data: Decomposes above 280 ºC	Estimated data: 312.76 ºC
Density		Experimental data: 1.769 at 50 ºC	Experimental results: 1.07-1.17 g/cm3 at 20 ºC
Vapour Pressure		Experimental data: 4.053E-07 Pa (partial pressure of ammonia over solid (NH4)2SO4 at 25 ºC)	Estimated data: 0.02 Pa at 25 ºC
Partition Coefficient (log Kow)		Experimental data: –5.1 at 25 ºC	Estimated data: -2.79
Water solubility		Experimental data: 764 g/L at 20 ºC	Experimental data: 1480 g/L at 4 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental data: In unsterilized soil, ammonium sulfate is mineralized fairly rapidly, and subsequently nitrified. Nitrification and de-nitrification processes also occur naturally in streams and rivers, as well as in many secondary sewage treatment processes	Experimental results on Ammonium Acetate, read-across from experimental data on Sodium Acetate and read-across from estimated data on Ammonia and Acetic Acid, based on functional group:   Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		Experimental data: (96 h) LC50 > 100 mg/L (Pimephales promelas)	Experimental data and read-across from Potassium Acetate, based on molecular weights:   LC50 = 392.70 mg/L.
Acute Toxicity to Aquatic Invertebrates		Experimental data: (96 h) LC50 > 100 mg/L (Asellus Intermedius, Daphnia magna, Dugesia tigrina and Gammarus fasciatus)	Read-across from experimental data on analogues Sodium Acetate, Potassium Acetate and Ammonia, based on molecular weights:   EC50 = 108.81 - 939.66 mg/L
Toxicity to Aquatic Plants		Experimental data: (18 d) EC50 = ca. 25476 mg/L (ca. 2700 mg NH3/L) (Chlorella vulgaris)	Read-across from experimental data on analogues Acetic Acid, Potassium Acetate and Ammonium Sulphate, based on molecular weights: (72 h) EC50 > 392.70 mg/L; (72 h) NOEC = 392.70 mg/L.
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LC50 >= 2000 mg/kg bw (rat, mouse) LC50 = 3040 mg/kg bw (mouse)	Weight of evidence: Read-across from experimental data on Potassium Acetate and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-3546.59 mg/kg bw
Acute Toxicity: Inhalation		Experimental data: (4 h) LC50 > 3.6 mg/m3 (rats) (1 h) LC50 > 2 mg/m3 (rabbits)	No data
Acute Toxicity: Dermal		Experimental data:   LC50 > 2000 mg/kg bw (rats, mice)	Weight of evidence: Read-across from experimental data on Fumaric Acid and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-26556.42 mg/kg bw
Skin Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits. -         Single exposure for 20 hours, intact skin: no skin effects observed. -         Multiple exposures, 8 hrs/day for 5 consecutive days, intact skin: no skin effects observed.	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate and Ammonium Lactate, and Ammonium Stearate based on functional group: The substance Ammonium Acetate is considered as not irritating for skin.
Eye Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits (50 mm3 of undiluted substance).	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate, Ammonium Sulphate, and Ammonium Stearate, based on functional group: The substance Ammonium Acetate is considered as not irritating for eyes.
Skin Sensitization		No data	Weight of evidence:   Read-across approach from experimental results on Citric Acid, Glycolic Acid, Sodium Glycolate, Lactic Acid, Ammonium Lactate, and Triacetin, based on functional group:   All this substances were not sensitising for human and guinea pigs. Based on these results, Ammonium acetate is considered to be not sensitizing.
Repeated Dose Toxicity		Experimental data: A 14-day inhalation study on rats exposed to 300 mg/m3, the only tested dose, did not report histopathological changes in the lower respiratory tract. As the respiratory tract is the target organ for inhalation exposure, the NOEL for toxicity to the lower respiratory tract is 300 mg/m3.   The NOAEL after feeding diets containing ammonium sulfate for 13 weeks to rats was 886 mg/kg bw/day. The only toxicity sign found was diarrhea in male animals of the high-dose group (LOAEL: 1792 mg/kg bw/day).	Repeated dose toxicity: oral: Weight of evidence: Experimental results:   Repeated dose toxicity: oral: 90 days withfemale Wistar rats. The NOAEL was 3150.4 mg/kg bw/day. Repeated dose toxicity: oral: 15 days study with female Wistar rats. The NOAEL 3102.2 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights:   The NOAEL >= 0.047 mg/kg bw/day, in male rats chronically treated for 8 months via drinking water. The NOAEL >= 3382.76 mg/kg bw/day, in male Wistar rats daily treated for 4 weeks by feed. The NOAEL >= 19.73 mg/kg bw/day, in male Long-Evans rats treated for 3 months in the diet. The NOAEL >= 0.0094 mg/kg bw/day, in male Wistar rats treated by drinking water for 112 days.    Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   The NOAEL >= 54 mg/kg bw/day, in albino rats treated for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	Experimental data: Ammonium sulfate was not mutagenic in bacteria (Ames test) and yeasts with and without metabolic activation systems.	Weight of evidence:   Read-across from Sodium Acetate (category analogue) based on functional group:   Reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with metabolic activation. Resultslead to the conclusion that Ammonium Acetate did not cause point mutations in the microbial systems. Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered to be not mutagenic on S.typhimurium TA 98, TA 100, TA 1535, TA 97, and/or TA 1537, with and without metabolic activation. Read-across from experimental data on Ammonia, anhydrous, based on functional group: Ammonium acetate is considered to be not mutagenic on Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA, with and without metabolic activation.   Read-across from experimental data on Ammonia, aqueous solution, based on functional group: Ammonium acetate is considered not mutagenic on E. coli Sd-4-73, without metabolic activation.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Ammonium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation. Read-across from the analogue Phenoxy acetic acid, based on functional group: Ammonium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation.   Estimated data from Danish (Q)SAR Database: Ammonium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	-         Chromosomal aberration	Experimental data:   Ammonium sulfate did not induce chromosomal aberrations in mammalian or human cell cultures.	Weight of evidence: Read-across from Sodium Acetate (category analogue) based on functional group:   In an in vitro chromosomal aberration assay with a Chinese hamster fibroblast cell line, CHL, without metabolic activation systems, it is concluded that Ammonium acetate did not induce chromosomal aberrations(including gaps). Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation. Read-across from Ammonium Sulfate, based on functional group: Ammonium Acetate is not considered mutagenic on Chinese Hamster Ovary cells, in the absence of a metabolic activation system.
Genetic Toxicity in vivo		No data	Key studies: Read-across from Sodium Acetate (category analogue) based on functional group:   The Testicular DNA-synthesis inhibition test (DSI test) on male mice provides evidence that Ammonium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Test substance did not inhibit DNA replication in this assay.
Carcinogenicity		Experimental data: Similarly to other salts, high doses of ammonium sulfate may have the capability of tumor promotion in the rat stomach; it is, however, much less potent than sodium chloride when tested under identical conditions.	No data
Reproductive Toxicity		TOXICITY TO REPRODUCTION: Fischer 344 rats (10 rats/sex/dose) were exposed during 13 weeks to diets containing 0, 0.38, 0.75, 1.5 or 3 % ammonium sulfate (corresponding to 0, 222, 441, 886, 1792 mg/kg bw/day in males and to 0, 239, 484, 961, 1975 mg/kg bw/day in females). No effects on the reproductive organs were observed. Therefore, the NOAEL was 886 mg/kg bw/day for males and 1975 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: No data.	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). A fertility test on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 722.25 mg/kg bw/day, and LOAEL greater than 722.25 mg/kg bw/day for reproductive effects. Read-across from the analogue Citric Acid, sodium salt, based on molecular weights: A fertility study on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 54.0 mg/kg bw/day, and LOAEL greater than 54.0 mg/kg bw/day for reproductive effects. Read-across from the analogue Ammonium sulfate, based on molecular weights: A study on male and female rats exposed for 13 weeks to diets with Ammonium Sulfate. For Ammonium Acetate, the NOAEL is calculated to be 1033.64 mg/kg bw/day for males, and 2304.12 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Experimental results: A study on female rats fed an ammonium-containing diet starting on day 1 of pregnancy until weaning (at posnatal day on 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. The NOAEL for developmental toxicity was 4293 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights: Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Ammonium Acetate, theNOAEL is calculated to be939.66 mg/kg bw/day (based on maternal toxicity: mortality, pregnancy and resorption; and on neonatal effects: mortality and body weight). Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). Read-across from the analogue substance Calcium Formate, based on molecular weights: A three-generation drinking water study was performed. For Ammonium Acetate, the NOAEL is calculated to be equal or higher than 236.96 mg/kg bw/day. Read-across from Acetic Acid, based on molecular weights: A one-generation study was performed on female mice, rats and rabbits with Acetic Acid. The read-across approach was applied and the NOAEL with the substance Ammonium acetate is calculated to be equal or greater than 2055.47 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw..

Executive summary:

Based on the experimental results (reported under the endpoint record 07.02.03_03 Ammonium sulfate) obtained with the analogue Ammonium sulfate (LD 50 for rats > 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

Reference 6

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The analogue Ammonium Sulfate which shares the same functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for acute dermal toxicity endpoint.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
See attached reporting format.

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

Read-across approach from published experimental data (study with a test method according to the Toxicity Guidelines of Japan (Ministry of Health and Welfare in Japan, 1984)) on the analogue Ammoniun sulphate.

GLP compliance:

not specified

Test type:

other: read-across from a fixed dose procedure with an analogue

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 333.28 mg/kg bw

Based on:

test mat.

Other findings:

Based on the experimental results obtained with the analogue Ammonium sulfate (LD 50 for mice > 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

The analogue Ammonium Sulfate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties for the aquatic toxicity endpoint. These properties are:
- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,
- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and
- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

The analogue Ammonium Sulphate, which shares the same ammonia/ammonium ion functional group with Ammonium Acetate, also has comparable values for the relevant molecular properties:

- a low log Pow value which is –5.1 at 25 ºC for Ammonium Sulphate and -2.79 for Ammonium Acetate,

- a similar water solubility which is 764 g/L at 20 ºC for Ammonium Sulphate and 1480 g/L at 4 ºC for Ammonium Acetate, and

- similar molecular weights which are 132.14 for Ammonium Sulphate and 77.08 for Ammonium Acetate.

As indicated in the European Chemical Agency Practical Guide 6 “How to report read –across and categories”, the structural grouping was realized using “OECD QSAR APPLICATION TOOL BOX” version 1.1.0. Presented results show that both substances have common (eco)toxicological behavior (attachment).

Table 1. Data Matrix, Analogue Approach.

CAS Number		Source chemical 7783-20-2	Target chemical 631-61-8
CHEMICAL NAME		Ammonium sulphate	Ammonium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Experimental data: Decomposes above 280 ºC	Experimental data: 114 ºC
Boiling Point		Experimental data: Decomposes above 280 ºC	Estimated data: 312.76 ºC
Density		Experimental data: 1.769 at 50 ºC	Experimental results: 1.07-1.17 g/cm3 at 20 ºC
Vapour Pressure		Experimental data: 4.053E-07 Pa (partial pressure of ammonia over solid (NH4)2SO4 at 25 ºC)	Estimated data: 0.02 Pa at 25 ºC
Partition Coefficient (log Kow)		Experimental data: –5.1 at 25 ºC	Estimated data: -2.79
Water solubility		Experimental data: 764 g/L at 20 ºC	Experimental data: 1480 g/L at 4 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental data: In unsterilized soil, ammonium sulfate is mineralized fairly rapidly, and subsequently nitrified. Nitrification and de-nitrification processes also occur naturally in streams and rivers, as well as in many secondary sewage treatment processes	Experimental results on Ammonium Acetate, read-across from experimental data on Sodium Acetate and read-across from estimated data on Ammonia and Acetic Acid, based on functional group:   Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		Experimental data: (96 h) LC50 > 100 mg/L (Pimephales promelas)	Experimental data and read-across from Potassium Acetate, based on molecular weights:   LC50 = 392.70 mg/L.
Acute Toxicity to Aquatic Invertebrates		Experimental data: (96 h) LC50 > 100 mg/L (Asellus Intermedius, Daphnia magna, Dugesia tigrina and Gammarus fasciatus)	Read-across from experimental data on analogues Sodium Acetate, Potassium Acetate and Ammonia, based on molecular weights:   EC50 = 108.81 - 939.66 mg/L
Toxicity to Aquatic Plants		Experimental data: (18 d) EC50 = ca. 25476 mg/L (ca. 2700 mg NH3/L) (Chlorella vulgaris)	Read-across from experimental data on analogues Acetic Acid, Potassium Acetate and Ammonium Sulphate, based on molecular weights: (72 h) EC50 > 392.70 mg/L; (72 h) NOEC = 392.70 mg/L.
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LC50 >= 2000 mg/kg bw (rat, mouse) LC50 = 3040 mg/kg bw (mouse)	Weight of evidence: Read-across from experimental data on Potassium Acetate and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-3546.59 mg/kg bw
Acute Toxicity: Inhalation		Experimental data: (4 h) LC50 > 3.6 mg/m3 (rats) (1 h) LC50 > 2 mg/m3 (rabbits)	No data
Acute Toxicity: Dermal		Experimental data:   LC50 > 2000 mg/kg bw (rats, mice)	Weight of evidence: Read-across from experimental data on Fumaric Acid and Ammonium Sulphate, based on molecular weights: LD50 = 2333.28-26556.42 mg/kg bw
Skin Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits. -         Single exposure for 20 hours, intact skin: no skin effects observed. -         Multiple exposures, 8 hrs/day for 5 consecutive days, intact skin: no skin effects observed.	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate and Ammonium Lactate, and Ammonium Stearate based on functional group: The substance Ammonium Acetate is considered as not irritating for skin.
Eye Irritation/Corrosion		Experimental data: This substance was not irritating for rabbits (50 mm3 of undiluted substance).	Weight of evidence: Read-across approach from experimental data on analogues Potassium Acetate, Ammonium Sulphate, and Ammonium Stearate, based on functional group: The substance Ammonium Acetate is considered as not irritating for eyes.
Skin Sensitization		No data	Weight of evidence:   Read-across approach from experimental results on Citric Acid, Glycolic Acid, Sodium Glycolate, Lactic Acid, Ammonium Lactate, and Triacetin, based on functional group:   All this substances were not sensitising for human and guinea pigs. Based on these results, Ammonium acetate is considered to be not sensitizing.
Repeated Dose Toxicity		Experimental data: A 14-day inhalation study on rats exposed to 300 mg/m3, the only tested dose, did not report histopathological changes in the lower respiratory tract. As the respiratory tract is the target organ for inhalation exposure, the NOEL for toxicity to the lower respiratory tract is 300 mg/m3.   The NOAEL after feeding diets containing ammonium sulfate for 13 weeks to rats was 886 mg/kg bw/day. The only toxicity sign found was diarrhea in male animals of the high-dose group (LOAEL: 1792 mg/kg bw/day).	Repeated dose toxicity: oral: Weight of evidence: Experimental results:   Repeated dose toxicity: oral: 90 days withfemale Wistar rats. The NOAEL was 3150.4 mg/kg bw/day. Repeated dose toxicity: oral: 15 days study with female Wistar rats. The NOAEL 3102.2 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights:   The NOAEL >= 0.047 mg/kg bw/day, in male rats chronically treated for 8 months via drinking water. The NOAEL >= 3382.76 mg/kg bw/day, in male Wistar rats daily treated for 4 weeks by feed. The NOAEL >= 19.73 mg/kg bw/day, in male Long-Evans rats treated for 3 months in the diet. The NOAEL >= 0.0094 mg/kg bw/day, in male Wistar rats treated by drinking water for 112 days.    Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   The NOAEL >= 54 mg/kg bw/day, in albino rats treated for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	Experimental data: Ammonium sulfate was not mutagenic in bacteria (Ames test) and yeasts with and without metabolic activation systems.	Weight of evidence:   Read-across from Sodium Acetate (category analogue) based on functional group:   Reverse mutation assay using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with metabolic activation. Resultslead to the conclusion that Ammonium Acetate did not cause point mutations in the microbial systems. Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered to be not mutagenic on S.typhimurium TA 98, TA 100, TA 1535, TA 97, and/or TA 1537, with and without metabolic activation. Read-across from experimental data on Ammonia, anhydrous, based on functional group: Ammonium acetate is considered to be not mutagenic on Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and TA 1538, and Escherichia coli WP2uvrA, with and without metabolic activation.   Read-across from experimental data on Ammonia, aqueous solution, based on functional group: Ammonium acetate is considered not mutagenic on E. coli Sd-4-73, without metabolic activation.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Ammonium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation. Read-across from the analogue Phenoxy acetic acid, based on functional group: Ammonium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation.   Estimated data from Danish (Q)SAR Database: Ammonium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	-         Chromosomal aberration	Experimental data:   Ammonium sulfate did not induce chromosomal aberrations in mammalian or human cell cultures.	Weight of evidence: Read-across from Sodium Acetate (category analogue) based on functional group:   In an in vitro chromosomal aberration assay with a Chinese hamster fibroblast cell line, CHL, without metabolic activation systems, it is concluded that Ammonium acetate did not induce chromosomal aberrations(including gaps). Read-across from Acetic Acid, based on functional group:   Ammonium Acetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation. Read-across from Ammonium Sulfate, based on functional group: Ammonium Acetate is not considered mutagenic on Chinese Hamster Ovary cells, in the absence of a metabolic activation system.
Genetic Toxicity in vivo		No data	Key studies: Read-across from Sodium Acetate (category analogue) based on functional group:   The Testicular DNA-synthesis inhibition test (DSI test) on male mice provides evidence that Ammonium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Test substance did not inhibit DNA replication in this assay.
Carcinogenicity		Experimental data: Similarly to other salts, high doses of ammonium sulfate may have the capability of tumor promotion in the rat stomach; it is, however, much less potent than sodium chloride when tested under identical conditions.	No data
Reproductive Toxicity		TOXICITY TO REPRODUCTION: Fischer 344 rats (10 rats/sex/dose) were exposed during 13 weeks to diets containing 0, 0.38, 0.75, 1.5 or 3 % ammonium sulfate (corresponding to 0, 222, 441, 886, 1792 mg/kg bw/day in males and to 0, 239, 484, 961, 1975 mg/kg bw/day in females). No effects on the reproductive organs were observed. Therefore, the NOAEL was 886 mg/kg bw/day for males and 1975 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: No data.	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). A fertility test on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 722.25 mg/kg bw/day, and LOAEL greater than 722.25 mg/kg bw/day for reproductive effects. Read-across from the analogue Citric Acid, sodium salt, based on molecular weights: A fertility study on female rats daily treated by feed for several months. For Ammonium Acetate, the NOAEL is calculated to be 54.0 mg/kg bw/day, and LOAEL greater than 54.0 mg/kg bw/day for reproductive effects. Read-across from the analogue Ammonium sulfate, based on molecular weights: A study on male and female rats exposed for 13 weeks to diets with Ammonium Sulfate. For Ammonium Acetate, the NOAEL is calculated to be 1033.64 mg/kg bw/day for males, and 2304.12 mg/kg bw/day for females.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Experimental results: A study on female rats fed an ammonium-containing diet starting on day 1 of pregnancy until weaning (at posnatal day on 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. The NOAEL for developmental toxicity was 4293 mg/kg bw/day . Read-across from the analogue Sodium Acetate, based on molecular weights: Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Ammonium Acetate, theNOAEL is calculated to be939.66 mg/kg bw/day (based on maternal toxicity: mortality, pregnancy and resorption; and on neonatal effects: mortality and body weight). Read-across from the analogue Citric Acid, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Ammonium Acetate, the NOAEL is calculated to be equal or greater than 3009.37 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). Read-across from the analogue substance Calcium Formate, based on molecular weights: A three-generation drinking water study was performed. For Ammonium Acetate, the NOAEL is calculated to be equal or higher than 236.96 mg/kg bw/day. Read-across from Acetic Acid, based on molecular weights: A one-generation study was performed on female mice, rats and rabbits with Acetic Acid. The read-across approach was applied and the NOAEL with the substance Ammonium acetate is calculated to be equal or greater than 2055.47 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw..

Executive summary:

Based on the experimental results (reported under the endpoint record 07.02.03_04 Ammonium sulfate) obtained with the analogue Ammonium sulfate (LD 50 for mice > 2000 mg/kg bw) and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

Endpoint conclusion

Dose descriptor:

LD50

Value:

2 338.28 mg/kg bw

Additional information

Acute oral toxicity:

Weight of evidence: Read-across approach from experimental results obtained with analogues Potassium Acetate and Ammonium Sulphate. All results were higher than 2000 mg/kg bw.

Read-across approach from Potassium Acetate data:

LD 50 for male rats = 3.25 (2.48-4.26) g/kg bw. Based on molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be 2.55 (1.95 -3.35) g/kg bw.

LD 50 for rats = 3250 mg/kg bw. Based on molecular weights, the read-across approach is applied and the LD50 for substance Ammonium Acetate is calculated to be 2552.58 mg/kg bw.

Read-across approach from Ammonium Sulphate data:

LD 50 for rats >= 2000 mg/kg bw. Based on molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be >= 2333.28 mg/kg bw.

LD 50 for mice > 2000 mg/kg bw. Based on molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw.

LD 50 for mice = 3040 (2670-3440) mg/kg bw. Based on molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be 3546.59 (3114.93-4013.25) mg/kg bw.

Acute dermal toxicity:

Weight of evidence: Read-across approach from experimental results obtained with analogues Fumaric Acid and Ammonium Sulphate. All results were higher than 2000 mg/kg bw.

Read-across approach from Fumaric Acid:

4h-LD 50 for New Zealand rabbits was higher than 20000 mg/kg bw. Based on these results and the molecular weights, the read-across approach is applied and the LD 50 for Ammonium acetate is calculated to be higher than 26556.42 mg/kg bw.

Read-across approach from Ammonium Sulphate:

The LD 50 for rats and mice was higher than 2000 mg/kg bw. Based on these results and the molecular weights, the read-across approach is applied and the LD50 for Ammonium Acetate is calculated to be > 2333.28 mg/kg bw for rats and mice.

Justification for classification or non-classification

Acute toxicity:

Oral: LD50 > 2000 mg/kg bw: non- classification

Dermal: LD50 > 2000 mg/kg bw: non- classification