Calcium di(acetate)

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive toxicity

Remarks:

based on generations indicated in Effect levels (migrated information)

Type of information:

migrated 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 Citric Acid which shares the same functional group with Calcium Acetate, also has comparable values for the relevant molecular properties for the toxicity to reproduction endpoint.

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

other: read-across

Title:

Unnamed

Year:

2010

Materials and methods

Principles of method if other than guideline:

Read-across approach from published experimental data on fertility and one-generation study on the analogue Citric acid.

GLP compliance:

no

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Citric acid

EC Number:

201-069-1

EC Name:

Citric acid

Cas Number:

77-92-9

IUPAC Name:

citric acid

Details on test material:

- Name of test material (as cited in study report): Citric acid
- Molecular formula (if other than submission substance): C6H8O7
- Molecular weight (if other than submission substance): 192.1
- Smiles notation (if other than submission substance): OC(=O)CC(O)(CC(O)=O)C(=O)O
- InChl (if other than submission substance): InChI=1/C6H8O7/c7-3(8)1-6(13,5(11)12)2-4(9)10/h13H,1-2H2,(H,7,8)(H,9,10)(H,11,12)
- Structural formula attached as image file (if other than submission substance): see Fig. in attached report

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not examined

Other effects:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Details on results (P0)

Based on the experimental results obtained with the analogue Citric acid (NOAEL >= 2500 mg/kg bw/day (basis for effect: number of pregnancies) in rats daily treated by feed before, during, and after mating), and the molecular weights, the read-across approach was applied and the NOAEL with the substance Calcium Acetate is calculated to be equal or greater than 3087.65 mg/kg bw/day.

The analogue Citric acid which shares the same functional group with Calcium Acetate, also has comparable values for the relevant molecular properties. These properties are:
- a low log Pow value, which is -1.72 for Citric acid, and -1.38 for Calcium acetate,
- a high water solubility, which is 1330 g/L for Citric acid, and 1000 g/L at 25 ºC for Calcium acetate, and
- similar molecular weights which are 192.1 for Citric acid, and 158.17 for Calcium acetate.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

>= 3 087.65 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: (number of pregnancies)

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Details on results (F1)

Based on the experimental results obtained with the analogue Citric acid (NOAEL >= 2500 mg/kg bw/day in rats (basis for effect: number of young born, or survival of young animals)), and the molecular weights, the read-across approach was applied and the NOAEL with the substance Calcium acetate is calculated to be equal or greater than 3087.65 mg/kg bw/day.

The analogue Citric acid which shares the same functional group with Calcium Acetate, also has comparable values for the relevant molecular properties. These properties are:
- a low log Pow value, which is -1.72 for Citric acid, and -1.38 for Calcium acetate,
- a high water solubility, which is 1330 g/L for Citric acid, and 1000 g/L at 25 ºC for Calcium acetate, and
- similar molecular weights which are 192.1 for Citric acid, and 158.17 for Calcium acetate.

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 3 087.65 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: (number of young born, or survival of young animals).

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

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 77-92-9	Target chemical 62-54-4
CHEMICAL NAME		Citric acid	Calcium acetate
PHYSICO-CHEMICAL DATA
Melting Point		Measured data: 153 ºC	Measured data: 160 ºC (decomposition)
Boiling Point		Decomposes	No data
Density		Measured data: 1.665 g/cm3 at 20 ºC	Experimental results: 1.5 kg/m3
Vapour Pressure		Estimated data: 5.6E-09 mm Hg	Estimated data: 0.73 Pa at 25 ºC
Partition Coefficient (log Kow)		Measured data: -1.72	Estimated data: -1.38
Water solubility		Measured data: 1330 g/L	Experimental results: 1000 g/L at 25 ºC
ENVIRONMENTAL FATE and PATHWAY
Aerobic Biodegradation		Experimental results: Readily biodegradable	Read-across from Sodium Acetate (category analogue): Readily biodegradable
ENVIRONMENTAL TOXICITY
Acute Toxicity to Fish		Experimental data: (96 h) LC 50 = 1516 mg/L	Weight of evidence:   Read-across from Sodium acetate (category analogue) based on molecular weights:   (96 h) LC 50 > 96.45 mg/L(Brachydanio rerio)     Read-across from Potassium acetate (category analogue) based on molecular weights:   (96 h) LC 50 > 402.92 mg/L (Oncorhynchus mykiss)   (96 h) LC 50 > 402.92 mg/L (Cyprinodon variegatus)   (96 h) LC 50 > 399.98 mg/L(Brachydanio rerio)
Acute Toxicity to Aquatic Invertebrates		Experimental data: (48 h) EC 50 = 1535 mg/L	Weight of evidence:   Read-across from Potassium acetate (category analogue) based on molecular weights:   (48 h) EC 50 > 370.28 mg/L(Daphnia magna) (48 h) EC 0 = 227.5 mg/L (Daphnia magna)   Read-across from Sodium acetate (category analogue) based on molecular weights:   (48 h) EC 50 > 964.09 mg/L(Daphnia magna)
Toxicity to Aquatic Plants		Experimental data: (72 h) EC 50 = 640 mg/L	Key studies: Read-across from Potassium acetate (category analogue) based on molecular weights:   (72 h) EC 50 > 402.92 mg/L (Skeletonema costatum) (72 h) NOEC = 402.92 mg/L (Skeletonema costatum)     Supporting studies: Read-across from the analogue substance Acetic Acid, based on molecular weights:   (8 d) Toxicity threshold (TT) = 5272.33 mg/L (Scenedesmus quadricauda)
MAMMALIAN TOXICITY
Acute Toxicity: Oral		Experimental data: LD 50 = 5790 – 7100 mg/kg bw (mice) LD 50 = 11700 mg/kg bw (rats)	Key study: Experimental results: LD50=2700 mg/kg bw LD50 = 1943 mg/kg bw (female rats) LD50 = 3160 - 4640 mg/kg bw (male rats)
Acute Toxicity: Inhalation		No data	Key study:   Experimental results:   (4 h) LC50 > 5.6 mg/L (rats)
Acute Toxicity: Dermal		Experimental data:   A 4-hour skin irritation study in rabbits exposed to a solution containing 60% citric acid caused erythema and edema. This study did not assess a lethal dose value. TheLD50 was not provided.	Key study:   Read-across from the analogue Fumaric acid, based on molecular weights: LD50 (4 h) > 27247.2 mg/kg bw (female New Zealand White rabbits)
Skin sensitisation		Experimental results:   Citric acid (2.5 % aqueous solution) is not sensitizing for the human skin.   No allergic reactions were seen when 60 patients with hand eczema, all of whom were involved in handling food, were patch tested (covered contact, probably 24 hr) with 2.5% citric acid in petrolatum.	Weight of evidence:   Read-across from the analogue substances Citric acid (source chemical), 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, Calcium acetate is also considered to be not sensitising.
Repeated Dose Toxicity		Repeated dose toxicity: oral: Experimental results:   In a 150-day study with male New Zealand White rabbits daily treated by feed, theNOAEL was 1500 mg/kg bw/day(based on no effects observed at the only dose tested).   In a 6-week study with male Sprague-Dawley rats daily treated by feed, theNOAEL was 4800 mg/kg bw/day(based on no effects observed at the highest dose tested).	Repeated dose toxicity: oral: Weight of evidence:   Read-across from Sodium acetate (category analogue), based on molecular weights:   Repeated dose toxicity: oral: 112-day study in male Wistar rats. The NOAEL was determined as 0.0096 mg/kg bw/day.   Repeated dose toxicity: oral: 3-month study in male Long-Evans rats. The NOAEL was determined as 20.25 mg/kg bw/day.   Repeated dose toxicity: oral: 4-week study in male Wistar rats. The NOAEL was determined as 3472.02 mg/kg bw/day.   Repeated dose toxicity: oral: 8-month study in male Long-Evans rats. The NOAEL was determined as 0.048 mg/kg bw/day.     Read-across from the analogue Citric acid, sodium salt, based on molecular weights:   TheNOAEL >= 55.41 mg/kg bw/day, in rats daily treated by feed for ca. 1 year.
Genetic Toxicity in vitro	-         Gene mutation in bacteria	Experimental data:   In a bacterial reverse mutation assays usingS. typhimurium(TA97, TA98, TA100 and TA104) in the presence and absence of metabolic activation and up to 2000μg/plate, citric 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 Calcium Acetate did not cause point mutations in the microbial systems. Read-across from the analogue substance Acetic Acid, based on functional group: Calcium 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.
	-         Mammalian gene mutation	No data	Weight of evidence: Read-across from the analogue Acetic anhydride, based on functional group: Calcium acetate is considered to be not mutagenic on mouse lymphoma L5178Y cells, with and without metabolic activation.   Read-across from the analogue Phenoxyacetic acid, based on functional group: Calcium acetate is considered to be not mutagenic on Chinese hamster ovary cells, with and without metabolic activation. Estimated data from Danish (Q)SAR Database: Calcium acetate was not mutagenic in mammalian cell gene mutation assays on mouse lymphoma L5178Y cells nor on Chinese hamster ovary cells.
	Chromosomal aberration	No data	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, Calcium acetate did not induce chromosomal aberrations(including gaps). Read-across from the analogue substance Acetic Acid, based on functional group: CalciumAcetate is considered as not clastogenic on Chinese hamster Ovary (CHO) cells, without metabolic activation.
Genetic Toxicity in vivo		Experimental data:   In a Dominant Lethal assay using male/female rats dosed at 3 g/kg for 5 days, citric acid did not induced germ cell genotoxicity.	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 Sodium acetate is not genotoxic in animals (basis of the method: measuring 3H-thymidine incorporation). Acetic acid, calcium salt did not inhibit DNA replication in this assay.
Carcinogenicity		No data	Data waiving (the substance is not classified as mutagen)
Reproductive Toxicity		TOXICITY TO REPRODUCTION: Experimental data: In a one-generation oral reproductive toxicity study, female rats and mice were daily treated with citric acid before, during, and after mating. The NOAEL was equal or greater than 2500 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young). In a fertility study, female rats were exposed to citric acid in their daily diet for several months. TheNOAEL (reproductive toxicity) was 600 mg/kg bw/day(based on no effects observed at the only dose tested).   DEVELOPMENTAL TOXICITY / TERATOGENICITY:   Experimental data:   In a one-generation oral reproductive toxicity study, female rats and mice were daily treated with citric acid before, during, and after mating. The NOAEL was equal or greater than 2500 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young).	TOXICITY TO REPRODUCTION: Weight of evidence: Read-across from the source chemical Citric Acid to the target chemical, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Calcium Acetate, the NOAEL is calculated to be equal or greater than 3087.65 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 Calcium Acetate, the NOAEL is calculated to be 741.04 mg/kg bw/day, and LOAEL greater than 741.04 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 Calcium Acetate, the NOAEL is calculated to be 55.41 mg/kg bw/day, and LOAEL greater than 55.41 mg/kg bw/day for reproductive effects.   DEVELOPMENTAL TOXICITY / TERATOGENICITY: Weight of evidence: Read-across from Sodium Acetate (category analogue) based on molecular weights:   Pregnant CD-1 mice were treated by oral gavage with Sodium Acetate on days 8-12 of gestation. For Calcium Acetate, theNOAEL is calculated to be equal or greater than 964.5 mg/kg bw/day for maternal toxicity and neonatal effects (mortality and body weight).   Read-across from the source chemical Citric Acid to the target chemical, based on molecular weights: A study on rats and mice daily treated by feed before, during, and after mating. For Calcium Acetate, the NOAEL is calculated to be equal or greater than 3087.65 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 Calcium Acetate, the NOAEL is calculated to be equal or higher than 243.13 mg/kg bw/day. Read-across from theanalogue substanceAcetic 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 Calcium acetate is calculated to be equal or greater than 2108.93 mg/kg bw/day for maternal and developmental toxicity in mice, rats, and rabbits.

Applicant's summary and conclusion

Conclusions:

The NOAEL with the substance Calcium acetate is calculated to be equal or greater than 3087.65 mg/kg bw/day (basis for effect: number of pregnancies, number of young born, or survival of young animals).

Executive summary:

Based on the experimental results (reported under the endpoint record 07.08.01_01 Citric acid) obtained with the analogue Citric acid (NOAEL >= 2500 mg/kg bw/day in rats (basis for effect: number of pregnancies, number of young born, or survival of young animals)), and the molecular weights, the read-across approach was applied and the NOAEL with the substance Calcium acetate is calculated to be equal or greater than 3087.65 mg/kg bw/day for studied effects.