calcium N2,N6-bis(3-carboxypropanoyl)lysine (1:1)

Endpoint summary

Description of key information

LD50 (oral) > 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January, from 14th to 28th, 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

December 17th, 2001

GLP compliance:

no

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy breeding, S. Pietro al Natisone (UD).
- Weight at study initiation: 192 - 202 g.
- Housing: transparent polycarbonate cages (425 x 266 x 180 mm); animals were housed 5 per cage.
- Fasting period before study: test animals were fasted overnight before the beginning of the test. 4 hours after exposure, regular pellet diet was provided.
- Diet: pellet diet from Harlan Italy.
- Water: purified water, ad libitum.
- Quarantine: 5 days.
- Health check: the animals were subjected to a veterinary examination to ensure their suitability for the study.

ENVIRONMENTAL CONDITIONS
- Illumination: fluorescent light lamp.
- Photoperiod: 12 hours light and 12 hours dark.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/ml

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 female

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: weekly weight control. Mortality, main organic functions evaluation, skin evaluation, mucous evaluation mobility and sensitivity evaluation were conducted at 30 min, at 2 and 4 hours and after 1, 2, 5, 6, 7, 8, 9, 12, 13 and 14 days of exposure.
- Necropsy of survivors performed: at the end of the study period.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed

Clinical signs:

During the test, all animals showed piloerection 4 hours after the exposure, tucked up abdomen 24 hours after the exposure.Those symptoms disappeared in two animals after 7 days from the test start, in three animals after 8 days from the test start.

Body weight:

A weight increase was observed in all test animals.

Other findings:

Two animals showed a moderate mucous enteritis and one animal showed a slight mucous enteritis.

Weight increase

Female N.	Weighed gain (g)
1	13
2	10
3	10
4	16
5	16

Body weight of treated animals, registered at the test start, after 7 and after 14 days of treatment.

Female N.	Start (g)	Mid period (g)	End (g)
1	197	199	210
2	202	203	212
3	200	203	210
4	195	198	211
5	192	195	208
X	197.2	199.6	210.2
DS	3.96	3.44	1.48

X= average

SD= standard deviation

Interpretation of results:

other: not classified, according to the CLP Regulation

Conclusions:

LD50 (rat female) > 2000 mg/kg bw

Executive summary:

Method

Acute oral toxicity study of the test item in Sprague Dawley rats was performed according to the OECD Guidelines No. 420 - Acute Oral Toxicity – fixed dose. In the present study, conducted as a ‘limit test’, single oral administration of the test item was made to a single group of five female Sprague Dawley rats at the dose of 2000 mg/kg bw, to assess its acute toxicity, through oral gavage.

After 4 hours from the administration, the normal pellet diet was provided. Following their treatment, the rats were observed for incidence of mortality and signs of toxicity for 14 days and then sacrificed and subjected to a necropsy examination. The rats were observed for: mortality, main organic functions evaluation, skin evaluation, mucous evaluation, mobility and sensitivity evaluation. The body weight was weekly recorded.

Observations

When tested at the dose of 2000 mg/kg bw, the test item did not induce any mortality in any of the treated rats. The body weight gained by treated rats was lower than species and strain standard during the first week of the study, and not found to be adversely affected during the second week. At gross necropsy, 2 animals showed a moderate mucous enteritis and 1 animal showed a slight enteritis. During the study, all animals showed piloerection 4 hours after treatment and tucked up abdomen after 24 hours from the administration. These symptoms disappeared in 2 animals after 7 days from the test start, and after 8 days in the other 3 tested animals.

Results

Based on these results, and according to the criteria for classification of chemicals OECD 420 the test article, is not classified under CLP regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

ACUTE ORAL TOXICITY

An acute oral toxicity study was conducted on the target chemical Set-Retard plus. The substance is expected to be biochemically stable and not bioavailable; it is expected to be mainly excreted as such, without interaction with the life-system. Because of the possibility of a partial metabolism/transformation of the substance cannot be excluded, the available information on possible precursors, which could be also the metabolites/transformation products of the target susbtance, has been precautionarly taken into account. The read across approach can be considered as reliable and appropriate for the endpoint under investigation; details on the read across approach are included in the reportattached to the IUCLID section 13.

Target Substance

In order to evaluate the acute toxicity by oral route of the target substance, an experimental study was conducted according to the OECD Guideline 420. During the test a single dose was administered by oral gavage to a five fimale group of Sprague Dawley rats (2000 mg/kg bw). Following their treatment, the rats were observed for incidence of mortality and signs of toxicity for 14 days and then sacrificed and subjected to a necropsy examination.

During the test no death occurred and no significative clinical signs were observed.

LD50 (rat female) > 2000 mg/kg bw

 

RA Substance 1a (succinic acid)

[Secondary source: OECD SIDS (2003) - Disodium Succinate CAS N°: 150-90-3. UNEP publications. Ref. KODAK, Company Reports]

Information are available from secondary source; unfortunately, only few details are reported. The substance was tested for acute oral exposure. No death observed at concentration of 2260 mg/kg bw, the effects observed are: weakness and diarrhea.

LD50 (rat male and female) > 2260 mg/kg bw

 

RA Substance 1b (monosodium succinate)

[Maekawa A. et al., (1990) Lack of toxicity/carcinogenicity of monosodium succinate in F344 rats; Food Chem. Toxicol. 28, 235 (1990)]

The acute oral toxicity of the substance, was examined in F344 rats. The test item was administered by gavage at dose of 500, 1000, 2000, 4000 and 8000 mg/kg bw.

At dose 8 g/kg one female rat died during the first week, all of the other rats survived until the end of the study. No clear toxicological effect was observed in rats that died or were killed, except for haemorrhage of the lung which was observed in some rats given the highest dose.

LD50 (rat male and female) > 8000 mg/kg bw

 

RA Substance 2b (L-lysine hydrochloride)

[Briglia R.J. et al., (1973) Effect of selected amino acids on ethanol toxicity in rats; Journal of Ppharmaceutical Sciences; Vol. 62, No.1, January 1973]

The substance has been tested for acute toxicity by oral route according to the Wilcoxon method. The rats were then subjected to the same neurological tests for signs of pharmacological effect.

LD50 (rat male and female): 10130 mg/kg bw

ACUTE INHALATION TOXICITY

Waiving – Exposure consideration

Taking into account the vapour pressure (VP = 0.00175 Pa) and the particle size distribution (10 % < 6.6 µm and 2.3 % < 2.0 µm) of the target substance, the exposure of humans via inhalation route is unlikely. Moreover, appropriate risk management measures and devices are implemented at the site(s) where the substance is manufactured and used, thus the possible occupational exposure and contact with the substance can be reduced at very low concentrations. General population can be considered not exposed to the substance.

 

ACUTE DERMAL TOXICITY

Waiving – Study scientifically unjustified

The physicochemical properties of the substance do not suggest potential for a significant rate of absorption through the skin. In the skin irritation test no adverse effects were seen and in the skin sensitisation assay no signs of systemic toxicity were recorded.

Furthermore, the substance is not acutely toxic by oral route, up to doses of 2000 mg/kg bw. Taking into account that the dermal absorption is commonly lower than the oral one, no systemic adverse effects are expected up to the same dose level by dermal route.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

In the case of the oral route, the acute toxicity hazard categories and acute toxicity estimates (ATE) defining the respective categories are:

- Category 1: ATE ≤ 5 mg/kg bw

- Category 2: 5 < ATE ≤ 50 mg/kg bw

- Category 3: 50 < ATE ≤ 300 mg/kg bw

- Category 4: 300 < ATE ≤ 2000 mg/kg bw

The acute oral LD50 in rats was established to be greater than 2000 mg/kg bw.

No data are available for acute dermal/inhalation toxicity, however no significant adverse effects are expected and no further investigations are required.

Therefore, the substance is not classified for oral acute toxicity, according to the CLP Regulation (EC 1272/2008).