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

Endpoint summary

Description of key information

Oral

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.

Dermal

Under the conditions of this study, the acute dermal LD50 value of the test material was found to be above 2 000 mg/kg bw in male and female CRL:(WI) rats.

Inhalation

According to Column 2 of REACH Annex VIII, in addition to the oral route, for substances other than gases, information shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure.  Exposure via the dermal route is more likely than via the inhalation route and data has therefore been provided for exposure via the dermal route.  Information on exposure via the inhalation route is not required.

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:

06 May 2003 to 06 June 2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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

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.

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.

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).

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Findings from this study was considered to be acceptable to address this endpoint. This non-GLP study was assigned a reliability score of 2 in line with criteria of Klimisch et. al. (1997).

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:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 May 2016 to 14 June 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15 to 25 ºC, below 70 RH %), protected from light.

Species:

rat

Strain:

other: CRL: (WI)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: Only specified as young adult.
- Weight at study initiation: 218 to 269 g
- Housing: Individual caging in Type II polypropylene/polycarbonate cages.
- Diet: Ad libitum; the food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Water: Tap water from the municipal supply, as for human consumption, from a 500 mL bottle, ad libitum.
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19.9 to 25.0 °C
- Humidity: 32 to 62 %
- Air changes: 15 to 20 air exchanges per hour
- Photoperiod: 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Back
- % coverage: 10 % (5 x 5 cm)
- Type of wrap if used: Sterile gauze pads (approximately 5 x 5 cm were placed on the skin of the rats to cover the dampened test material. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing: At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied: 2000 mg/kg bw
- For solids, paste formed: The test material was dampened with sufficient water before application to ensure good contact with the skin.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 animals per sex per dose.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test material and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

- Necropsy of survivors performed: Yes
Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital. After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

- Other examinations performed: The body weights were recorded on Day 0 (before test material administration) and on Days 7 and 14 just before necropsy.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The test material did not cause mortality at the dose level of 2000 mg/kg bw.

Clinical signs:

other: There were no systemic clinical signs noted in any animal throughout the study. No local dermal signs were observed after treatment with the test material during the 14 days observation period.

Gross pathology:

There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the acute dermal LD50 value of the test material was found to be > 2000 mg/kg bw in male and female CRL:(WI) rats.

Executive summary:

An acute dermal toxicity study was performed with test material in CRL:(WI) rats, in accordance with the standardised guideline OECD Guideline No.: 402, under GLP conditions.

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test material was applied as a single dermal 24-hour exposure followed by a 14-day observation period. The test material was applied to the shaven backs of the animals to cover approximately 10 % of the total body surface area, it was applied on a gauze pad kept in contact with the skin using an adhesive patch. The trunk of the animal was then wrapped with semi-occlusive plastic wrap for 24 hours.

Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

The test material did not cause any mortality at 2000 mg/kg bw, there were no systemic clinical or local dermal signs observed throughout the study. Body weight gains of the treated animals showed no indication of a test-material related effect. There were no macroscopic findings to report. 

Under the conditions of this study, the acute dermal LD50 value of the test material was found to be above 2000 mg/kg bw in male and female CRL:(WI) rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

One study is available to address this endpoint. The study was performed in accordance with standardised guidelines and under GLP conditions. The study was assigned a reliability score of 1 in line with the principles for assessing data quality as defined by Klimisch et al. (1997). The quality of the database is therefore considered to be good.

Additional information

Oral

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”. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

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.

Dermal

An acute dermal toxicity study was performed with test material in CRL:(WI) rats, in accordance with the standardised guideline OECD Guideline No. 402, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test material was applied as a single dermal 24-hour exposure followed by a 14-day observation period. The test material was applied to the shaven backs of the animals to cover approximately 10 % of the total body surface area, it was applied on a gauze pad kept in contact with the skin using an adhesive patch. The trunk of the animal was then wrapped with semi-occlusive plastic wrap for 24 hours.

Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

The test material did not cause any mortality at 2 000 mg/kg bw, there were no systemic clinical or local dermal signs observed throughout the study. Body weight gains of the treated animals showed no indication of a test-material related effect. There were no macroscopic findings to report. 

Under the conditions of this study, the acute dermal LD50 value of the test material was found to be above 2000 mg/kg bw in male and female CRL:(WI) rats.

Inhalation

In accordance with section 8.5.2 of Column 2 of REACH Annex VIII, in addition to the oral route, for substances other than gases, information shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. Exposure via the dermal route is more likely than via the inhalation route and data has therefore been provided for exposure via the dermal route. Information on exposure via the inhalation route is not required.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to acute toxicity via the oral or dermal routes.