Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Feb - Mar 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
Dec 17, 2001
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
(from the competent authority) Landesanstalt für Umwelt Baden-Württemberg
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Reference substance name:
Reaction mass of diammonium (2R,3R)-2,3-dihydroxybutanedioate and diammonium bis[(2R,3R)-2,3-dioxidobutanedioato]diantimonate(2-)
EC Number:
943-693-3
IUPAC Name:
Reaction mass of diammonium (2R,3R)-2,3-dihydroxybutanedioate and diammonium bis[(2R,3R)-2,3-dioxidobutanedioato]diantimonate(2-)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: 170492F01
- Expiration date of the Batch: June 22, 2019
- Content:
21.4 g/100 g Test Item
23.5 g/100 g (NH4)2C4H4O6
30 g/100 g Tartrate
6.7 g/100 g Ammonium
9.1 g/100 g Antimony
53.5 g/100 g Water
- Physical state / color: liquid / colorless to yellowish, clear
- Density: 1.305 g/mL

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Solubility and stability of the test substance in the solvent/vehicle: The stability of the test item in the vehicle was determined indirectly by concentration control analysis.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item preparation for each test group was produced shortly before administration by stirring with a magnetic stirrer. The homogeneity of the test item preparations during administration was ensured by stirring with a magnetic stirrer.

FORM AS APPLIED IN THE TEST (if different from that of starting material): For the high dose, the liquid test item was administered unchanged. For the lower doses, an administration volume of 3 mL/kg bw of suitable test item preparations was used to facilitate administration.

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl:WI (Han) SPF
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: young adult animals, approx. 10 weeks
- Weight at study initiation: animals of comparable weight (+/- 20 % of the mean weight)
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: single housing in fully air-conditioned rooms
- Diet (e.g. ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: at least 5 days

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

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: unchanged (high dose) or deionized water (mid and low dose)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: undiluted (high dose), 22.0 g/100 mL (mid dose), 3.7 g/100 mL (low dose)
- Justification for choice of vehicle: solution in deionized water

MAXIMUM DOSE VOLUME APPLIED: 3 mL/kg bw

DOSAGE PREPARATION (if unusual): The test item preparation for each test group (doses of 300 and 50 mg a.i./kg bw) was produced shortly before administration by stirring with a magnetic stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: By request of the sponsor a starting dose of 50 mg a.i./kg bw was chosen in the first step with 3 female animals. Because no mortality occurred in this step, a further dose of 300 mg a.i./kg bw was administered to another group of 3 female animals in the second step. As no animal died, a further dose of 2000 mg a.i./kg bw was administered to 3 female rats in the third step. All animals died in this step. Therefore, a further dose of 300 mg a.i./kg bw was administered to another group of 3 female animals in the fourth step. Due to a purity of approx. 45 % the doses were increased to 4444 mg/kg bw to ensure a dose of 2000 mg a.i./kg bw, 667 mg/kg bw to ensure a dose of 300 mg a.i./kg bw and 111 mg/kg bw to ensure a dose of 50 mg a.i./kg bw, respectively.
Doses:
4444 mg/kg bw (high dose), 667 mg/kg bw (mid dose), 111 mg/kg bw (low dose)
The doses were regarding to the test item concentration of approx. 45 % increased to ensure an actual dose of 2000, 300 or 50 mg a.i./kg bw for the main components.
No. of animals per sex per dose:
3 female animals per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the day of death on study day 1. Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter. A check for any dead or moribund animals was made at least once each workday.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Statistics:
Calculations were performed using Microsoft Excel 2010 and checked with a calculator.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
act. ingr.
Mortality:
In the 2000 mg a.i./kg bw test group all animals died within 1 or 2 hours or day 1 after administration, respectively.
No mortality occurred in both 300 mg a.i./kg bw test groups and in the single 50 mg a.i./kg bw test group.
Clinical signs:
In the 2000 mg a.i./kg bw test group one animal showed impaired general state and piloerection at hour 1 after administration.
In other animal of this test group impaired general state was seen from hour 1 until hour 3, while poor general state was from hour 4 until hour 5. Furthermore, piloerection was observed from hour 1 until hour 5, while dyspnoea was seen from hour 2 until hour 5. In addition, cowering position was seen from hour 4 until hour 5 and diarrhea was observed from hour 2 until hour 3 after administration.
The third animal showed a good clinical condition at hour 0, but died at hour 1.
In all animals of the first 300 mg a.i./kg bw test group impaired general state and piloerection were seen from hour 1 until hour 5 after administration, while dyspnoea was seen in these animals from hour 1 until hour 3 or 5. In one animal of this test group, diarrhea was seen at hour 2.
In all animals of the second 300 mg a.i./kg bw test group impaired general state and piloerection were seen from hour 1 until hour 5 after administration.
In all animals of the single 50 mg a.i./kg bw test group impaired general state and piloerection were seen from hour 3 until hour 5 after administration.
Body weight:
All animals gained weight in a normal range throughout the study period.
Gross pathology:
The following macroscopic pathologic findings were observed in the animals that died in the 2000 mg a.i./kg bw test group:
- Edema in the lung lobes in one animal
- Dark, spotted discoloration of all lung lobes in two animals
- Dark, spotted discoloration of the liver in all animals
- Liquid, brownish discolored stomach contents in all animals
- Dark red discoloration of the glandular stomach in one animal
- Red discoloration of the small intestine in two animals
There were no macroscopic pathological findings in any animal sacrificed at the end of observation period (300 mg a.i./kg bw: 6 females, 50 mg a.i./kg bw: 3 females).

Any other information on results incl. tables

Table 1: Mortality

Dose [mg a.i./kg bw]

Sex

Administration

No. of animals

Mortality [animals]

2000

Female

1

3

3

300

1

3

No mortality

300

2

3

No mortality

50

1

3

No mortality

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Under the conditions of this study the median lethal dose of the test substance after oral administration was found to be greater than 300 mg a.i./kg bw and less than 2000 mg a.i./kg bw in rats.
Executive summary:

In an acute oral toxicity study performed according to the Acute Toxic Class Method, doses of 2000, 300 and 50 mg active ingredient (a.i.)/kg bw of the test item (undiluted or preparations in deionized water) were administered by gavage to four test groups of three fasted Wistar rats each (2000 mg a.i./kg bw in 3 females, 300 mg a.i./kg bw in 6 females and 50 mg a.i./kg bw in 3 females).

Since the test item contained approx. 55 % water, the amount was adjusted to the purity of approx. 45 %, i.e. the dose was increased to 4444 mg/kg bw to ensure an actual dose of 2000 mg a.i./kg bw, 667 mg/kg bw to ensure an actual dose of 300 mg a.i./kg bw and 111 mg/kg bw to ensure an actual dose of 50 mg a.i./kg bw.

The following test substance related clinical observations were recorded, clinical signs occurred within the first day after administration:

2000 mg a.i./kg bw (first test grpup):

- Mortality in all animals

- Impaired general state in two animals

- Poor general state in one animal

- Dyspnoea in one animal

- Piloerection in two animals

- Cowering position in one animal

- Diarrhea in one animal

- Macroscopic pathological findings in the animals that died: edema in the lung lobes in one animal; dark, spotted discoloration of all lung lobes in two animals; dark, spotted discoloration of the liver in all animals; liquid, brownish discolored stomach contents in all animals; dark red discoloration of the glandular stomach in one animal; red discoloration of the small intestine in two animals

300 mg a.i./kg bw (first test group):

- No mortality occurred

- Impaired general state in all animals

- Dyspnoea in all animals

- Piloerection in all animals

- Diarrhea in one animal

300 mg a.i./kg bw (second test group):

- No mortality occurred

- Impaired general state in all animals

- Piloerection in all animals

50 mg a.i./kg bw (single test group):

- No mortality occurred

- Impaired general state in all animals

- Piloerection in all animals

All animals gained weight in a normal range throughout the study period.

There were no macroscopic pathological findings in any animal sacrificed at the end of the observation period (300 mg a.i./kg bw: 6 females, 50 mg a.i./kg bw: 3 females).

The acute oral LD50 was calculated to be > 300 < 2000 mg a.i./kg bw.