Reaction mass of copper digluconate, sodium sulphate and copper sulphate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04.Apr.07 - 30.Jul.07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

OECD Guideline for the Testing of Chemicals, Guideline 420 Acute Oral Toxicity, Fixed Dose Procedure, updated 17 December 2001

Deviations:

yes

Remarks:

The animals received A04C diet instead of TEKLAD 2014 as was indicated in the Study Plan. This had no ímpact on the Study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Version / remarks:

Commission Directiva 2004/73/EC of 29 April 2004. Annex 28. Method 8.1 bis

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Commission Directiva 2001/59/EC of 6 August 2001, Annex VI

GLP compliance:

yes

Test type:

acute toxic class method

Test material

Test material information

Constituent 1

Reference substance name:

Copper di-D-gluconate

EC Number:

208-408-2

EC Name:

Copper di-D-gluconate

Cas Number:

527-09-3

Molecular formula:

C12H22CuO14

IUPAC Name:

copper(2+) bis(2,3,4,5,6-pentahydroxyhexanoate)

Constituent 2

Reference substance name:

Sodium sulphate

EC Number:

231-820-9

EC Name:

Sodium sulphate

Cas Number:

7757-82-6

Molecular formula:

H2O4S.2Na

IUPAC Name:

disodium sulfate

Constituent 3

Reference substance name:

Copper sulphate

EC Number:

231-847-6

EC Name:

Copper sulphate

Cas Number:

7758-98-7

Molecular formula:

CuSO4

IUPAC Name:

copper(2+) ion sulfate

Test material form:

liquid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (range of 20 ± 5 ce) and in the dark.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The necessary amount of test item was weighed in a volumetric flask and distilled water was added gradually until the desired volume was reached.
The formulation was later transferred to an adequate container and kept in agitation. The necessary volume of vehicle was added to the formulation prepared on 3 May 2007 in the final container to make up the required volume. The vehicle added was first poured into the volumetric flask before being added to the final container.

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Wistar Hannover HsdRccHan: WIST rat

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Ibérica, S. L. {Ctra. Sant Miquel del Fai, km 3, 08182- Sant Feliu de Codines, Barcelona, Spain)
- Number of animals: 6 females
- Age when treated: 8-9 weeks
- Weight at study initiation: 158-164 g

CONDITIONS
Standard Laboratory Condltions.: Air-conditioned with target ranges for room temperature at 19-26 °C, relative humidity of 40-70% and 10-20 air changes per hour. Room temperature and relative humidity were monitored continuously. Values outside these ranges
occasionally occurred. These transient variations are not considered to have any influence on the study and therefore these data are not reported but are retained at RCC CIDA S.A. There was a 12-hour fluorescent light/12-hour dark cycle.

ACCOMMODATION
Sighting study: Individual in Makrolon type-5 cages {59.0 x 38.5 x 20.0 cm) with sawdust bedding {supplied by Harlan lnterfauna Ibérica, S.L.).
Main study: Groups of four or five in Makrolon type-5 cages {59.0 x 38.5 x 20.0 cm) with Lignocel 3-4 sawdust bedding (supplied by Harlan lnterfauna Ibérica, S.L.).

- Diet: Pelleted standard SAFE A04C rat/mouse maintenance diet (Scientific Animal Food & Engineering, Route de St. Bris, 89290-Augy, France, (Batch 70206}, Expiry date: 6 February 2008) ad libitum. Results of analyses for contaminants are archived at RCC CIDA S.A.
- Water: Tap water in bottles ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC CIDA S.A.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Distilled water (Aqua B. Braun, Ref. 387875, Batches:7083B06, 7154B07

Doses:

lnitially, one rat was treated at the dose of 300 mg/kg. 8ecause no signs of toxicity were recorded in thís animal, another animal was treated at the dose of 2000 mg/kg

No. of animals per sex per dose:

One group of five females Wistar Hannover HsdRccHan rats (including one used in the initial sighting study)

Control animals:

not specified

Statistics:

No statistical analysis was used.

Results and discussion

Preliminary study:

The animal treated at 2000 mg/kg is reported under "9.2 MAl N STUDY'.
No mortality nor clinical signs were observed in the animal treated at 300 mg/kg during the course of the study.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

One animal died at the dosage level of 2000 mg/kg approximately one hour after administration.

Clinical signs:

other: The animal that died after administration showed decreased motor activity, abnormal gait, hunched back, decreased muscle tone, prostration, clonic convulsions, dyspnea and diarrhea during approximately 30 minutes following administration and before dying.

Gross pathology:

Right renal pelvis slightly dilated was recorded in one animal. No macroscopic alterations were recorded at the necropsies performed in the others animals that survived the treatment.
Hemorrhagic area in cranial surface of 0.5 x 0.5 cm diameter approximately and presence of product in the stomach were observed in the animal that died after the administration.

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

DL50> 2000 mg/kg. Substance is not classified according CLP criteria.

Executive summary:

In the main study, one group of five females Wistar Hannover HsdRccHan rats (including one used in the initial sighting study) were treated with the test item by oral gavage administration at a dosage of 2000mg/kg body weight. The test item was diluted in vehicle (distilled water) at a concentration of 0,2g/mL and administered at a dosing volume of 10mL/ kg.

During the sighting study, one additional animal was dosed at 300mg/ kg without showing any signs of toxicity.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2 -15. Mortality/viability was recorded at approximately 30minutes, 1, 2,3 and 5 hours after weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

The death of one of the animals treated at the dose of 2000mg/ kg, was recorded approximately one hour after the administration.

Among the animals treated at this dose hunched back and abnormal gait were recorded on the day of adminsitration.

The naimls thata died after the adminsitration also had decreased motor activity, abnormal gait, hunched back, decreased muscle tone, prostration, clonic convulsions, dyspnea and diarrhea.

Two of the animals thata survived the treatment showed huncled back, abnormal gait, pallor, dark red urine, dark feces andyellowish skin and mucus in the days following tretament. In one of the animals, these clinical signs were recorded at the most until day 6 after adminsitration. The other animal showed abnormal gait until day 9, and the hunched back until day 11 after adminsitration. Another animal showed abnormal gait and dark feces one day after administration, and pallor on days 3 and 4 after adminsitration.

No clinical signs were recorded from day 12 onwards.

Hemorrhagic area in cranial surface of 0.5 x 0.5 cm diameter and presence of product in stomach, were observed in the animal that died after administration.

In one animal was recorded right renal slightly diated. No macroscopical alterations were recorded at the necropsies performed in the others animals that survived the treatment.

The body weight of the animals was within the range commoly recorded for this strain and age.