Bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-04-18 to 2017-05-25

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)

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

EC Number:

300-491-4

EC Name:

Bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

Cas Number:

93940-93-3

Molecular formula:

C10H20CuN2O6

IUPAC Name:

bis[N-(2-hydroxyethyl)-N-methylglycinato-N,O,ON]copper

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Remarks:

WISTAR rats Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

Housing and Feeding Conditions:
- Full barrier in an air-conditioned room
- Temperature: 22 ± 3 °C
- Relative humidity: 55 ± 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x per hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua ad injectionem

Details on oral exposure:

The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.

Doses:

The starting dose was selected to be 300 mg/kg body weight (step 1 and step 2)
step 3 performed with 2000 mg/kg body weight

No. of animals per sex per dose:

3 + 3 females step 1 and 2
3 females step 3

Control animals:

no

Details on study design:

- Observation Period:
The surviving animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.

- Weight Assessment:
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

- Clinical Examination:
A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

- Pathology:
At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally at a dosage of 250-400 mg/kg bw.
All animals were subjected to gross necropsy and examined macroscopically for gross pathological changes. In the absence of gross pathological changes no tissues were preserved for a possible histopathological evaluation.

- Evaluation of Results:
Results were interpreted according to OECD Guideline 423, Annex 2 and GHS.
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.
With few exceptions, data were captured using the validated departmental computerised system E WorkBook (version 10.1.2, ID Business Solutions Ltd.).

Statistics:

n.a.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

ca. 500 mg/kg bw

Based on:

test mat.

Mortality:

Two animals treated with the test item at a dose of 300 mg/kg bw (step 1 and 2) died spontaneously within 3 days post dosing.
All animals treated with the test item at a dose of 2000 mg/kg bw (step 3) died spontaneously within 30 - 60 minutes post dosing.

Clinical signs:

The most relevant clinical findings in the animals treated with the test item at a dose of 300 mg/kg bw were reduced spontaneous activity, sunken flunks, piloerection, half eyelid closure, slow movements and hunched posture.

The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, ataxia, half eyelid closure, abnormal breathing, prone position, sunken flunks and opisthotonus.

Body weight:

300 mg/kg bw: Throughout the 14-day observation period, the weight gain of the surviving animals was within the normal range of variation for this strain.

Gross pathology:

At necropsy, no macroscopic findings were observed in any animal of any step.

Any other information on results incl. tables

Results per Step

Step	Sex / No.	Starting Dose (mg/kg bw)	Number of Animals	Number of Intercurrent Deaths
1	Female / 1 - 3	300	3	1
2	Female / 4 - 6	300	3	1
3	Female / 7 - 9	2000	3	3

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The median lethal dose of the test item after a single oral administration to female rats, observed over a period of 14 days is: LD50 cut-off (rat): 500 mg/kg bw.

Executive summary:

The purpose of this study was to assess the toxicity of the test item when administered to rats as a single oral dose.

The study was carried out in accordance with OECD Guideline No. 423 and EPA Health Effects Test Guideline OPPTS 870.1100. The study was performed according to GLP regulations.

Two groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 300 mg/kg body weight. The test item was suspended with aqua ad injectionem (sterile water) at a concentration of 0.03 g/mL and administered at a dose volume of 10 mL/kg.

One group of three female WISTAR Crl: WI(Han) rats, was treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was suspended with aqua ad injectionem (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

Two animals treated with the test item at a dose of 300 mg/kg bw (step 1 and 2) died spontaneously within 3 days post dosing. All remaining animals survived until the end of the study. The most relevant clinical findings in the animals treated with the test item at a dose of 300 mg/kg bw were reduced spontaneous activity, sunken flunks, piloerection, half eyelid closure, slow movements and hunched posture. Throughout the 14-day observation period, the weight gain of the surviving animals was within the normal range of variation for this strain.

All animals treated with the test item at a dose of 2000 mg/kg bw (step 3) died spontaneously within 30 - 60 minutes post dosing. The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, ataxia, half eyelid closure, abnormal breathing, prone position, sunken flunks and opisthotonus. At necropsy, no macroscopic findings were observed in any animal of any step.

The median lethal dose of the test item after a single oral administration to female rats, observed over a period of 14 days is: LD₅₀cut-off (rat): 500 mg/ kg bw.