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Description of key information

Acute oral toxicity

In an acute oral toxicity study in female CRL:(WI) SPF rats, following the acute toxic class method according to OECD guideline 423, the LD50 was observed to be between 300 and 2000 mg/kg bw in female CRL:(WI) rats (referring to anhydrous zirconium dinitrate oxide) (Matting, 2015a, scored Klimisch 1). According to the GHS and CLP criteria, zirconium dinitrate oxide can be classified as 'Category 4' for acute oral toxicity.

Acute inhalation toxicity

No reliable studies have been identified. Endpoint coverage is not required since the endpoints on acute oral toxicity as well as acute dermal toxicity are covered. Moreover, the inhalation route of exposure is less relevant as zirconium dinitrate oxide is manufactured and used as aqueous solutions.

Acute dermal toxicity

In an acute dermal toxicity study in CRL:(WI) SPF rats (5 males and 5 females), following the fixed dose procedure according to OECD guideline 402, the LD50 was observed to be above 2000 mg/kg bw (referring to hydrated zirconium oxynitrate) (Bioassay, 2018, scored Klimisch 1). No mortality was observed. As a result, the substance does not need to be classified for acute dermal toxicity.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014-10-29 to 2014-11-20
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)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: CRL:(WI) SPF
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
- Age at dosing: young, healthy adult rats, 9-10 weeks old
- Weight at treatment: 208-233 g
- Fasting period before study: yes, the night before treatment, food but not water was withheld during an overnight period. Food was returned 3 hours after treatment.
- Housing: 3 animals per cage, in Type II polypropylene/polycarbonate cages with Lignocel Bedding for laboratory animals
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.9-25.0°C
- Humidity (%): 30-70%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2014-10-29 To: 2014-11-20
Route of administration:
oral: gavage
Vehicle:
other: PBS (phosphate buffered saline)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 or 200 mg/mL (referring to anhydrous zirconium dinitrate oxide)
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: The selection of the vehicle was made during trial formulation with the test item in distilled water and PBS at 3 concentrations that could be used for dosing 2000, 300 and 50 mg/kg as active ingredient (anhydrous zirconium dinitrate oxide). PBS was used further in the study, since the test item has an extremely low pH in distilled water.
- Lot/batch no. (if required): BCBJ5262V
- Purity: no data
- Expiry date: 2017-02-28

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION (if unusual):
The test item was freshly formulated at a concentration of 200 or 30 mg/mL (as active ingredient, anhydrous zirconium dinitrate oxide) in the vehicle, in the Central Dispensary Unit of CiToxLAB Hungary Ltd. on the day of administration. The formulation container was stirred continuously up to finishing the treatment.
CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The initial dose level was most likely to produce mortality in some of the dosed animals. In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose.
Doses:
2000 mg/kg bw in one group (as active ingredient) (anhydrous zirconium dinitrate oxide)
300 mg/kg bw in two groups (as active ingredient) (anhydrous zirconium dinitrate oxide)
No. of animals per sex per dose:
3 females per group
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 all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual observations were performed on skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor acitvity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter. Body weight of the animals found dead was recorded at necropsy
- Necropsy of survivors performed: yes. Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia. After examination of external appearance, the cranial, thoracic and the abdominal cavities were opened and organs and tisses were observed. Macroscopic abnormalities were recorded.



Statistics:
no data
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
act. ingr.
Remarks:
anhydrous zirconium dinitrate oxide
Mortality:
Mortality was observed in all animals receiving a single dose of 2000 mg/kg bw (3/3) on Day 1 or 2 or 4. No mortality was observed at the dose level of 300 mg/kg bw.
Clinical signs:
At 2000 mg/kg bw (expressed as anhydrous zirconium dinitrate oxide), the test item caused decreased activity (3/3), hunched back (3/3), incoordination (2/3), piloerection (2/3), dyspnoea (1/3), red discharge from the eye and around the nose (1/3).
At 300 mg/kg bw, hunched back was observed in 5/6 animals. All animals were symptom-free 6 hours after the treatment until the end of the observation period.
Body weight:
Body weight gains of test item treated animals during the study showed no indication of a test item-related effect at the dose level of 300 mg/kg bw.
Gross pathology:
- In animals found dead (from 2000 mg/kg bw group): Diffuse red coloration of the glandular stomach mucosa found in 2/3 found dead females, was considered to be potentially test item-related. However, definitive attribution of this change to the test item cannot be made in absence of histopathological examination. The changes in the lungs were typical for found dead animals.
- In surviving animals: No macroscopic observations were seen at a dose level of 300 mg/kg bw.
Other findings:
- Deviation from the study plan: Due to a typographical error, the pH value of the test item was reported to be 1.73 for the 7.35 mg/mL concentration in distilled water and 1.21 for the 294 mg/mL concentration in PBS in the Study plan. The correct values are 1.79 and 0.68, respectively. This deviation is considered to have no impact on the outcome of the study and interpretation of the results.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Under the conditions of this study, the acute oral LD50 value of zirconium dinitrate oxide was found to be between 300 and 2000 mg/kg bw in female CRL:(WI) rats. According to the GHS and CLP criteria, zirconium dinitrate oxide can be ranked as Category 4 for acute oral exposure.
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: adult rats
- Weight at study initiation: between 200 and 300 g
- Fasting period before study: no data
- Housing: maintained in air-conditioned rooms
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
50% aqueous solutions were used
Doses:
Single dose, no more data
No. of animals per sex per dose:
24 rats in total
Control animals:
no
Details on study design:
- Duration of observation period following administration: 10 days
Statistics:
The LD50 values were obtained from ten day mortality data by using the log-probability method.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 500 mg/kg bw
Based on:
test mat.
Mortality:
The rats showed a progressive depression and decrease in activity until death occurred.
No sex differences were noted, and the LD50 value was therefore derived from the combined data on both sexes.
The time of death varied from a few hours to a few days after administration. Few deaths however occurred later than 5 days after administration.
Clinical signs:
No characteristic physiologic changes were observed.
Body weight:
No characteristic physiologic changes were observed.
Gross pathology:
No characteristic gross pathologic changes were observed.
Conclusions:
The acute oral LD50 value in rats administered a 50% zirconyl nitrate aqueous solution via oral gavage is 2500 mg/kg bw (test item-based).
Endpoint conclusion
Endpoint conclusion:
adverse effect observed

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
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:
Nov 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Material tested is a substance related to the reference substance which is considered sufficiently similar to be covered as well by the dossier of the reference substance. This merging is also agreed by the SIEF and is reflected in the substance identity section of the dossier (Section 1.1). Therefore, even though the CAS numbers might differ, the tested material sufficiently represents the reference substance. Therefore, provision of a read across justification is not required.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
February 24, 1987
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
(from the competent authority) Landesanstalt für Umwelt Baden-Württemberg
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: room temperature
- Solubility and stability of the test substance in the solvent/vehicle: The homogeneity of the test item preparation during application was ensured by stirring with a magnetic stirrer. The stability of the test item in the vehicle was determined indirectly by concentration control analysis. For this purpose, the samples taken were stored at room temperature over the maximum duration of the administration period, subsequently deep-frozen and sent to the sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was ground with a mortar and pestle. The test item preparation was produced shortly before applied by stirring with a high-speed homogeniser (Ultra-Turrax) and a magnetic stirrer.

FORM AS APPLIED IN THE TEST (if different from that of starting material)
Suspension
Species:
rat
Strain:
Wistar
Remarks:
Crl:WI (Han) SPF
Sex:
male/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: males approx. 8 weeks, females approx. 12 weeks
- Weight at study initiation: animals of comparable weight (± 20 % of the mean weight)
- Housing: single housing in Makrolon cage type III, 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 before the beginning of the experimental phase

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
Type of coverage:
semiocclusive
Vehicle:
corn oil
Remarks:
Ph. Eur.
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal and dorsolateral parts of the trunk, about 40 cm²
- % coverage: at least 10 % of the body surface
- Type of wrap if used: semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): warm water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 mL/kg bw
- Concentration (if solution): 40 g/100 mL
- Constant volume or concentration used: yes
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
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 application (day 0), weekly thereafter and on the last day of observation. Clinical signs for each animal were recorded several times on the day of application and at least once during each workday thereafter.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, mortality, pathology
Statistics:
Calculations were performed using Microsoft Excel 2010 and checked with a calculator.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality occurred
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination. No local skin effects were observed.
Body weight:
The body weight of all male animals and of two female animals increased within the normal range throughout the study period. Two other female animals showed slight loss of body weight during the first week, while another animal showed stagnation of body weight within this week. All three animals gained weight in a normal range during the second week. Due to the fact that stagnation or slight loss of body weight is commonly known for females after dermal application, this stagnation is considered to be unspecific.
Gross pathology:
No macroscopic pathologic abnormalities were noted in any animal examined on the last day of observation (5 males and 5 females).

Table 1: Mortality

Dose [mg/kg bw]

2000

2000

Sex

Male

Female

Application

1

1

No. of animals

5

5

Mortality (animals)

No mortality

No mortality

Table 2: Maximum incidence of systemic clinical signs

Dose [mg/kg bw]

2000

Sex

Male

Female

Application

1

No. of animals

10

Animal No.

R 658

R 659

R 660

R 661

R 662

R 663

R 664

R 665

R 666

R 667

Abnormalities

-

-

-

-

-

-

-

-

-

-

Table 3: Nature and duration of local clinical signs

Dose [mg/kg bw]

2000

Sex

Male

Female

Application

1

No. of animals

10

Animal No.

R 658

R 659

R 660

R 661

R 662

R 663

R 664

R 665

R 666

R 667

Abnormalities

-

-

-

-

-

-

-

-

-

-

Table 4: Individual body weight changes

Dose [mg/kg bw]

2000

Sex

Male

Female

Application

1

Animal No.

R 658

R 659

R 660

R 661

R 662

R 663

R 664

R 665

R 666

R 667

Body weight at study day [g]:

 

 

 

 

 

 

 

 

 

 

0

225

217

222

224

217

210

209

207

205

194

7

261

255

260

252

252

206

203

208

215

199

14

299

288

301

286

285

216

210

216

221

207

Table 5: Gross pathology

Dose [mg/kg bw]

2000

Sex

Male

Female

Application

1

No. of animals

10

Animal No.

R 658

R 659

R 660

R 661

R 662

R 663

R 664

R 665

R 666

R 667

Abnormalities

-

-

-

-

-

-

-

-

-

-

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study the median lethal dose (LD50) of the test substance after dermal application was found to be greater than 2000 mg/kg bw in male and female rats. No mortality was observed.
Executive summary:

In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the test substance (as suspension in corn oil Ph. Eur.). The clipped application site (dorsal and dorso-lateral parts of the trunk, comprising at least 10 % of the total body surface) was covered by semi-occlusive dressing during the 24-hour exposure period. The animals were observed for 14 days.

- No mortality occurred.

- Neither signs of systemic toxicity nor local skin effects were observed.

- The body weight of all male animals and of two out of five female animals increased within the normal range throughout the study period. Two other female animals showed slight loss of body weight during the first week, while another animal showed stagnation of body weight within this week. All three females gained weight in a normal range during the second week. Due to the fact that stagnation or slight loss of body weight is commonly known for females after dermal applications, this stagnation is considered to be unspecific.

- No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study (5 males and 5 females).

Accordingly, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw for male and female rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Acute oral toxicity

The single dose oral toxicity of zirconium dinitrate oxide was investigated according to the acute toxic class method (OECD guideline 423) in CRL:(WI) rats (Klimisch 1, Matting, 2015a). This study was assigned as key study. One group of 3 female CRL:(WI) rats was treated with the test item at a dose level of 2000 mg/kg bw (doses referring to anhydrous zirconium dinitrate oxide). Two groups of 3 female CRL:(WI) rats were treated with the test item at a dose level of 300 mg/kg bw.

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was administered formulated in PBS (phosphate buffered saline) at a concentration of 200 or 30 mg/mL at a dose volume of 10 mL/kg bw. Initially, 3 females were treated at a dose level of 2000 mg/kg bw. The test item caused mortality (3/3) in this group; therefore next 3 animals were treated at a dose level of 300 mg/kg bw. The test item did not cause mortality in this group; therefore a confirmatory group was treated with 300 mg/kg bw. No mortality was observed in this confirmatory group, therefore no further testing was required. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

Mortality was observed in all animals receiving a single dose of zirconium dinitrate oxide at 2000 mg/kg bw (3/3) on day 1, 2 or 4. No mortality was observed at the dose level of 300 mg/kg bw. At 2000 mg/kg bw, the test item caused decreased activity (3/3), hunched back (3/3), incoordination (2/3), piloerection (2/3), dyspnoea (1/3), red discharge from the eye and around the nose (1/3). At 300 mg/kg bw, hunched back was observed in 5/6 animals. All animals were symptom-free from 6 hours after the treatment until the end of the observation period. Body weight gains of test item treated animals during the study showed no indication of a test item-related effect at the dose level of 300 mg/kg bw. Diffuse red discoloration of the glandular stomach mucosa, found in 2/3 found dead females from the 2000 mg/kg bw group, was considered to be potentially test item-related. However, definitive attribution of this change to the test item cannot be made in absence of histopathological examination. The changes in the lungs were typical for found dead animals. In surviving animals, no macroscopic observations were seen at a dose level of 300 mg/kg bw.

Under the conditions of this study, the acute oral LD50 was found to be between 300 and 2000 mg/kg bw in female CRL:(WI) rats (referring to anhydrous zirconium dinitrate oxide).

In addition, in a supporting study, an acute oral LD50 value of 2500 mg/kg bw was observed for a 50% zirconyl nitrate aqueous solution (test item-based) (Klimisch 2, Cochran et al., 1950).

Acute inhalation toxicity

No reliable data available. No data required.

Acute dermal toxicity

In an acute dermal toxicity study performed following a Fixed Dose Procedure in agreement with OECD guideline 402 and GLP principles (Bioassay, 2018), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of zirconium oxynitrate hydrate (CAS 14985-18-3), the hydrated form of zirconium dinitrate oxide (CAS 13826-66-9), as suspension in corn oil Ph. Eur. The clipped application site (dorsal and dorso-lateral parts of the trunk, comprising at least 10% of the total body surface) was covered by semi-occlusive dressing during the 24-hour exposure period. The animals were observed for 14 days.

- No mortality occurred.

- Neither signs of systemic toxicity nor local skin effects were observed.

- The body weight of all male animals and of two out of five female animals increased within the normal range throughout the study period. Two other female animals showed slight loss of body weight during the first week, while another animal showed stagnation of body weight within this week. All three females gained weight in a normal range during the second week. Due to the fact that stagnation or slight loss of body weight is commonly known for females after dermal applications, this stagnation is considered to be unspecific.

- No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study (5 males and 5 females).

 

Accordingly, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw for male and female rats.

This study was considered reliable without restrictions (Klimisch 1) and was considered to be the key study to cover the acute dermal toxicity endpoint.

Acute toxicity: other routes

In the study of Cochran et al. (1950), an LD50 of 1250 mg/kg bw (test item-based, test item = 50% zirconyl nitrate aqueous solution) was obtained after intraperitoneal administration. This was considered as supporting information.

Justification for classification or non-classification

According to the CLP criteria and based on the available data, zirconium dinitrate oxide is classified for Acute Toxicity Category 4. However, no classification is required for acute toxicity after dermal exposure.

No data are available to decide on the classification for acute toxicity after inhalation exposure.