Reaction mass of isomers of disodium amino-(3-carboxylatopropanamido)hexanoate and trisodium 2,6-bis(3-carboxylatopropanamido)hexanoate

Endpoint summary

Description of key information

In the acute oral toxicity study with the test item in rats the determined LD50 is greater than 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:

August 2, 2016 - August 31, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

17 December 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

30 May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10 weeks old in group 1 and group 2
- Weight at study initiation: 236-237 g (group 1) and 215-234 g (group 2)
- Fasting period before study: The day before treatment the animals were fasted.
- Housing: 3 animals/sex/cage, Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets
- Diet: ad libitum, ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany
- Water: ad libitum, tap water
- Acclimation period: 20 days in first group and 21 days in second group

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua purificata Ph.Hg. VIII.

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Justification for choice of vehicle: Test item is soluble in vehicle.
- Lot/batch no.: 1602-5519

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: The starting dose was selected on the basis of the available information about the test item.
The method was conducted using a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 animals per dose (3 per step)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day.
Animals were observed individually after dosing once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h, after the treatment and once per day for 14 days thereafter. The body weight were recorded on day 0 (shortly before the treatment), on day 7 and on day 15 on all animals with a precision of 1 g, respectively.
- Necropsy of survivors performed: yes

Statistics:

none

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The test item did not induce mortality at a dose of 2000 mg/kg bw (group 1 and 2, step 1 and 2). All rats in step 1 and step 2 survived until the end of the 14-day observation period.

Clinical signs:

other: No treatment related symptoms were observed in the 2000 mg/kg bw test item dose groups (group 1 and 2, step 1 and 2) throughout the 14-day post-treatment period.

Gross pathology:

All animals treated with 2000 mg/kg bw of the test item survived until the scheduled necropsy on Day 15. Severe hydrometra was found in one female of the group 2. Hydrometra is a physiological finding and connected to the cycle of the animal. No pathological changes were found related to the test item during the macroscopic examination of animals.

Interpretation of results:

GHS criteria not met

Conclusions:

In the acute oral toxicity study with the test item in rats the determined LD50 is greater than 2000 mg/kg bw.

Executive summary:

An acute oral toxicity study was carried out using the class method according to OECD guideline 423. The method was conducted using a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. The starting dose was selected on the basis of the available information about the test item.

No animal died in the first step at 2000 mg/kg bw dose level, so three further female rats were treated with the same (2000 mg/kg bw) dose. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 were met.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on the 15th day after the treatment.

Lethality, Clinical symptoms and Body weight:

No lethality was noted following oral administration of a single dose of 2000 mg/kg bw dose groups.

No clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of all experimental animals were normal.

The body weight development was normal in all animals.

Gross pathology:

All animals survived until the scheduled autopsy on Day 15. All organs of all animals proved to be free of gross pathological changes.

Evaluation:

The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study with the test item in rats the determined LD50 is above 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant study

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:

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
Due to the low vapour pressure of the test item, which was calculated to be 1.24E-5 Pa at 20°C and 1.78E-5 Pa at 25°C, no exposure via inhalation is expected. Therefore, a study in acute inhalation toxicity can be omitted.

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:

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
Testing for acute dermal toxicity is not necessary if (i) the substance is not classified for oral acute toxicity and (ii) if in in vivo studies using dermal application (skin sensitization) no systemic effects were observed. As the substance is not classified for oral toxicity and in the LLNA assay no systemic effects were reported a study on dermal acute toxicity is not required.

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral toxicity

An acute oral toxicity study was carried out using the class method according to OECD TG 423. The method was conducted using a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. The starting dose was selected on the basis of the available information about the test item.

No animal died in the first step at 2000 mg/kg bw dose level, so three further female rats were treated with the same (2000 mg/kg bw) dose. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD TG 423 were met.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on the 15th day after the treatment.

Lethality, Clinical symptoms and Body weight:

No lethality was noted following oral administration of a single dose of 2000 mg/kg bw dose groups.

No clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of all experimental animals were normal.

The body weight development was normal in all animals.

Gross pathology:

All animals survived until the scheduled autopsy on Day 15. All organs of all animals proved to be free of gross pathological changes.

Evaluation:

The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study with the test item in rats the determined LD50 is above 2000 mg/kg bw.

Justification for classification or non-classification

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

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on acute toxicity, the test item is not classified for acute oral toxicity according to Regulation (EC) No 1272/2008, as amended for the seventeenth time in Regulation (EU) 2021/849.