Copper, diamminebis(nitrato-kO)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

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

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

without significant impact on the study

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Copper, diamminebis(nitrato-kO)

EC Number:

604-942-9

Cas Number:

15414-04-7

Molecular formula:

CuH6N4O6

IUPAC Name:

Copper, diamminebis(nitrato-kO)

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

The female rats were nulliparous and non-gravid, 8-10 weeks old at the beginning of the study.

Acclimation period: 5 days. Rats were initially housed 2 per cage, and then transferred to individual cage before dosing start at day 0. The animals’ health was assessed by a clinical observation, performed on the day of arrival at the facility, and daily during the acclimation period. During the acclimation period, the animals were housed in the study room.

Environment conditions: the animal room conditions were set as follows: temperature of 20 to 24°C, relative humidity of 40 to 70%, lighting of 12h / 24 h, ventilation of 10 to 20 cycles/hour of filtered, non-recycled air.

Housing: the animals were housed in polysulfone cages (type 3 high) containing autoclaved dust-free bedding. Cages were changed at least once a week. Each cage was labelled with the study number, the lot number, the cage number, the type of treatment and the animals’ identification (identification number and sex).

Food: specific rat food was dispensed ad libitum to the animals during the study in stainless troughs, except during the fasting period (food was withheld over-night before treatment to 3-4 hours after treatment).

Water: all animals had free access to tap water, contained in polycarbonate bottles supplied with stainless steel teats.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Milli-Q water

Details on oral exposure:

The animals were fasted the day before dosing. The access to food was restored 3 hours after the treatment. Free access to drinking water was maintained throughout the fasting period. The volume to administer to each animal was calculated based on the body weight measured on the morning of dosing. MNP363 volume was collected from the preparation flask, while continuously stirring (appendix 2). This volume was administered by gavage using a rigid cannula, on the morning of day 0 (D0).
The administration of the higher dose was carried because the rats treated with the initial dose survived.

Doses:

The initial dose of 300 mg/kg was chosen according to the OECD n°423 guideline and a higher dose (2000 mg/kg) was subsequently selected based on the absence of mortality observed in the rats treated with the initial dose.

No. of animals per sex per dose:

3 female rats: initial dose of 300 mg/kg
3 female rats: initial dose (confirmation) of 300 mg/kg
3 female rats: second dose of 2000 mg/kg

Control animals:

no

Details on study design:

Morbidity and mortality were assessed at least daily. The date of euthanasia and the animals’ body weight at that time were recorded, and a necropsy was conducted.

Health status was monitored via clinical observations. Animals were individually observed according to the following schedule: once on the day of arrival, once daily during the acclimation period, several times on the day of treatment (D0), daily from D1 to D14 (once or twice per day). Each abnormal symptom was noted as well as the time of appearance, duration and time of disappearance.

Individual body weights were reported according to the following schedule: once on the day of arrival, once on the day of selection, once on day -1 (before fasting period), once on day 0, before treatment, once daily from D1 to D14.

Animals were euthanized by an intra-peritoneal injection of a lethal dose of pentobarbital in a separate room than the necropsy room in order to decrease the animal stress. Euthanized animals were introduced in the necropsy room and a necropsy was performed. It consisted of the macroscopic observation of the external surfaces, all orifices, the external surfaces of the brain, the cranial, thoracic, abdominal and pelvic cavities, and the neck with their associated organs and tissues. Macroscopic abnormalities observed on these organs were recorded according to the facility standard operating procedures.

Statistics:

Not performed

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No unscheduled death was observed on rats treated with the test item at 300 mg/kg during the observation period. Two rats out of three died the day of the treatment with the test item at the dose of 2000 mg/kg.

Clinical signs:

No clinical sign of toxicity was observed on rats in the 300 mg/kg group, during the observation period. Several clinical signs of toxicity were observed on rats in the 2000 mg/kg group, during the observation period. Two rats showed successively prostration, reduced motor activity, ventral decubitus, piloerection, eyelids partially closed, coloured eyes outline red, excessive salivation, abdominal respiration between the 10 min and 3 hours following treatment. One other rat showed successively prostration, reduced motor activity, ventral decubitus, piloerection and eyelids partially closed between 10 min and 45 min post-treatment. These effects were reversed 6 hours after treatment excepted for piloerection which was reversed 6 days after treatment.

Body weight:

At reception, all individual body weights were within ± 20% of the average body weight of the previously dosed animals. It was also the case at D0 and throughout the in-life observation period. The body weight gain was similar in all animals.

Gross pathology:

In the 300 mg/kg dosed rats and one 2000 mg/kg dosed rat, macroscopic observations indicated normal external surfaces, all orifices, the external surfaces of the brain, the cranial, thoracic, abdominal and pelvic cavities, and the neck with their associated organs and tissues.
In two 2000 mg/kg dosed rats, macroscopic observations indicated coloured eyes outline red, excessive salivation, brown stains on nose, soiled area around the anus and discoloration (blue) of the digestive tract in one rat and discoloration (blue) of the digestive tract and mucosal redness in the jejunum and ileum in the other rat.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

According to our data, in the protocol based on the OECD n°423 guideline, MNP363 induced the dead of two rats out of three at the dose of 2000 mg/kg, and was nontoxic at the dose of 300 mg/kg in two consecutive sessions of three rats. According to these findings and to the EC No. 1272/2008 Regulation, the test item MNP363 is classified in category 4 with the hazard statement “H302, Harmful if swallowed”.