Bis(5-oxo-L-prolinato-N1,O2)zinc

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 May 2003 to 06 June 2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The test guideline states that the age of the rats should be 8 to 12 weeks. In this reports the rats were 6 weeks old at the start of administration. This is a deviation but should not invalidate the results.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Bis(5-oxo-L-prolinato-N1,O2)zinc

EC Number:

239-473-5

EC Name:

Bis(5-oxo-L-prolinato-N1,O2)zinc

Cas Number:

15454-75-8

Molecular formula:

C10H12N2O6Zn

IUPAC Name:

(3'aR)-3,3',6,6'-tetraoxo-3a,3'a,4,4',5,5',6,6'-octahydro-3H,3'H-1,1'-spirobi[pyrrolo[1,2-c]1-oxa-3-aza-2-zincacyclopentane]-1,1-bis(ylium)-2,2'-diuide

Test material form:

other: solid

Test animals

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Males: 6 weeks, females: 6 weeks
- Weight at study initiation: Males: 181.5 to 184.8 g, females: 143.5 to 146.4 g
- Fasting period before study: All animals was fasted for approximately 20 hours, from the day before administration until observation of acute toxicity symptoms was complete.
- Housing: Metal bracket cages, 1 animal per cage.
- Diet: γ-ray sterilized solid food for test animals (CRF-1) ad libitum.
- Acclimation period: 6 days during which the following observations and tests were implemented, and judgments were made as to whether the animals should be used in the study.
Clinical signs: A general observation of clinical signs was carried out once daily for 6 days in the morning.
Body weight measurement: Body weights of all of the animals were measured at the beginning and the end of quarantine/acclimation period.
- Water: Tap water ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 3 °C
- Humidity (%): 50 ± 20 %
- Air changes (per hr): All-fresh ventilation of 10 to 15 cycles per hour
- Photoperiod (hrs dark / hrs light): 12 hours, from 7 A.M. until 7 P.M.; between 200 and 600 lux.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

TEST MATERIAL ADMINISTRATION:
Animals recieved a single dose of the test material by oral administration. Oral administration of a volume of 10 mL per 1 kg of body weight was carried out using disposable syringes and oral catheters for rats. The administration of the test material was carried out once. All animals was fasted for approximately 18 hours before administration.

DOSAGE PREPARATION
3.0 g of the test article was directly weighed in a 20-mL graduated cylinder, after which distilled water for injection was added, the preparation fluid inside the graduated cylinder was transferred out and in again, and the mixture was thoroughly agitated. Then, the solution was transferred to a 15-mL graduated cylinder and prepared by diluting it to 15 mL with injection solvent.
Concentration of sample to be administered: 20 % (W/V).

- Rationale for the selection of the starting dose:
Since it was predicted based on toxicity information of similar products that toxicity would be low, the dosage was set to 2.0 g/kg, which is the maximum dosage prescribed in the OECD Guideline for the Testing of Chemicals, Number 423.

Doses:

2.0 g/kg

No. of animals per sex per dose:

3 female rats and 3 male rats per group.

Control animals:

no

Details on study design:

- Duration of observation period following administration: On the day of administration (Day 1), observation of acute toxicity symptoms was carried out for all of the surviving animals until approximately 2 hours after administration. In addition, observation of clinical signs was carried out for all of the surviving animals before administration and 6 hours after administration on Day 1, as well as once a day in the morning on Days 2 to 15.
- Frequency of observations and weighing: Body weights of all of the animals were measured at the beginning and the end of quarantine/acclimation period. The body weight of all remaining animals was measured on Days 1, 2, 6, 10, and 15.
- Necropsy of survivors performed: Yes. Necropsy examinations were carried out upon the death of a male rat (00202) and female rat (50203) that died midway through the study.
All surviving animals were sacrificed humanely by exsanguinating through the abdominal aorta under deep ether anesthesia 14 days after administration (Day 15). After a visual examination, macroscopic observations of various organs such as the abdominal cavity, pleural cavity, and cephalic region were carried out.

Statistics:

No statical analysis was performed.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

During the observation period, the death of 1 male rat on the second day after administration, and the death of 1 female rat on the third day after administration were confirmed.

Clinical signs:

other: During the observation period for acute toxicity symptoms, decrease in locomotor activity, lying on side, salivation, eyes closed (half or fully), lack of strength, and soft stools were observed. Decrease in locomotor activity, soft stools, and hypothermi

Gross pathology:

No abnormalities were confirmed in the surviving animals. For the animals that died, all of the findings shown in the table were similar for both sexes, and therefore it was considered that these changes were due to administration of the test article. The findings suggest that the test article severely irritates the mucous membranes of the gastrointestinal tract (especially the gastric mucosa).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, it was estimated that the LD50 value of the test material is greater than 2.0 g/kg body weight when dosed to male and female SD rats.

Executive summary:

The acute oral toxicity of the test material was investigated in a study which was conducted in accordance with the standardised guideline OECD Guideline for the Testing of Chemicals, Number 423 “Acute Oral Toxicity-Acute Toxic Class Method”.

During the study, the animals were stratified based on their body weight and randomly sorted into groups of 3 rats, with each group having a uniform average body weight. The animals received a single dose of the test material by oral administration at a dose of 2.0 g/kg bw and a dosage volume of 10 mL per kg of body weight. Administration was carried out using disposable syringes and oral catheters for rats. The vehicle used in this study was distilled water.

During the observation period for acute toxicity symptoms, decrease in locomotor activity, lying on side, salivation, eyes closed (half or fully), lack of strength, and soft stools were observed. Decrease in locomotor activity, soft stools, and hypothermia were observed six hours following administration. On the day following administration, diarrhoea was observed in 1 male rat; hypothermia and diarrhoea were observed in 1 female rat; decrease in locomotor activity, hypothermia, and diarrhoea were observed in another female rat. From among these 3 rats, the one in which a decrease in locomotor activity was observed the day following administration died on Day 3. All clinical signs had disappeared by Day 3 in the surviving animals.

Under the conditions of this study, it was estimated that the LD50 value of the test material is greater than 2.0 g/kg body weight when dosed to male and female SD rats.