trisodium (2S)-2,6-bis(3-carboxylatopropanamido)hexanoate

Endpoint summary

Description of key information

The substance was assessed for oral acute toxicity. In the oral toxicity study, concentrations up to 2000mg/kg bw were tested and the LD50 oral was determined being 5000 mg/kg bw.

Thus, the substance is not subject to classification for acute oral toxicity according to CLP.

Data for acute inhalation toxicity are not required as the substance is not used as aerosol and due to the low vapour pressure of -0.184 Pa no relevant vapour exposure is possible.

No dermal study as the substance does not meet the criteria for classification as acute toxicity by oral route and no systemic effects have been observed in the available skin irritation study.

Based on acute oral toxicity test results, the substance was shown not to be subject to classification according to CLP (Regulation EC No. 1272/2008).

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:

2019

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)

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 23ADB/50
- Expiration date of the lot/batch: 21/11/2023

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
In order to get the test item in a solution or suspension, which is applicable to the animals, several vehicles (sterile water, corn oil) were evaluated and preparation and solubility tests were performed. The test material formed an emulsion in corn oil which could not be applied. The preparation with sterile water yielded an applicable solution and considered to be adequate.

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 – 9 weeks
- Weight at study initiation: animal no. 1: 173 g; animal no. 2: 184 g; animal no. 3: 153 g; animal no. 4: 176 g; animal no. 5: 170 g
- Fasting period before study: n/a
- Housing: Full barrier in an air-conditioned room
- Diet: ad libitum
- Water : ad libitum
- Acclimation period: Adequate acclimatisation period (at least five days) under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22  3 °C
- Humidity (%): 55  10%
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): Artificial light, sequence being 12 hours light, 12 hours dark

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua ad injectionem (sterile water, DELTAMEDICA, lot no. 807634, expiry date: 06/2021). This vehicle was chosen due to its non-toxic characteristics.

Details on oral exposure:

Animal No. /
Sex Dose(mg/kg bw) Body Weight on Day of Administration (g) Volume of Test Item Preparation in Vehicle Administered per Animal (mL)
1 / Female 2000 173 1.7
2 / Female 2000 184 1.8
3 / Female 2000 153 1.5
4 / Female 2000 176 1.8
5 / Female 2000 170 1.7

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 animals, Fixed dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were weighed on day 1 (prior to the administration) and on days 8 and 15. A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). Thereafter, the animals were observed for clinical signs once daily until the end of the observation period.
- Necropsy of survivors performed: yes
- Other examinations performed: Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
All animals were subjected to gross necropsy and examined macroscopically for gross pathological changes. In the absence of gross pathological changes no tissues were preserved for a possible histopathological evaluation.

Preliminary study:

Sighting Study
The starting dose for the sighting study was 2000 mg/kg body weight.
No compound-related mortality was recorded for the first animal dosed.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

none

Clinical signs:

other: none

Gross pathology:

No specific gross pathological changes were recorded for any animal.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present study, a single oral application of the test item Trisodium (2S)-2,6-bis(3-carboxylatopropanamido) hexanoate_Disuccinyl lysine sodium salt to rats at a dose of 2000 mg/kg body weight was not associated with signs of toxicity or mortality.
The median lethal dose of Trisodium (2S)-2,6-bis(3-carboxylatopropanamido) hexanoate_Disuccinyl lysine sodium salt after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): ˃ 5000 mg/kg bw

Executive summary:

Five females WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was dissolved in the vehicle aqua ad injectionem (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

Table 1: Results

Animal No. / Sex	Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths
Sighting Study
1 / Female	2000	1	0
Main Study
2 / Female	2000	1	0
3 / Female		1	0
4 / Female		1	0
5 / Female		1	0

 

All animals survived until the end of the study without showing any signs of toxicity.Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain. At necropsy, no treatment-related macroscopic findings were observed in any animal of any step.

LD50cut-off (rat):                     ˃5000 mg/kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Reason / purpose for cross-reference:

data waiving: supporting information

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The substance was tested for oral acute toxicity in and in-vivo study.

Based on the findings in this study, the substance is not classified according to CLP (Regulation EC No. 1272/2008).