reaction mass of copper sulfate and deta to Reaction products of 2,2'-iminodi(ethylamine) and copper sulphate (1:1)

Endpoint summary

Description of key information

No study was available for the substance itself but the result for a category member was used.

The LD50 of the test item Reaction product of copper sulfate and triethylenetetramine is higher than 50 mg/kg body weight and lower than 300 mg/kg by oral route in the rat. In accordance with the O.E.C.D. Test Guideline No. 423, the LD50 cut-off of the test item may be considered as 300 mg/kg body weight by oral route in the rat.

According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with the Regulation EC No. 1272/2008, the test item Reaction product of copper sulfate and triethylenetetramine has to be classified in category 3. The signal word “Danger” and hazard statement H301 “Toxic if swallowed” are required.

The same classification was used for Copper diethylenetriamine sulfate.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH

1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
See the documentation for the category approach.

2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL
See the documentation for the category approach.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 17th, 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle – France)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 or 9 weeks
- Weight at study initiation: 183 - 228 g
- Fasting period before study: no
- Housing:
Healthy female rats were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet and water (e.g. ad libitum):
Drinking water (tap-water from public distribution system) and foodstuff (ENVIGO - 2016) were supplied ad libitum. Food was removed on day 1 and then redistributed 4 hours after the test item administration.
Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE). The results of analysis were kept in the Quality Assurance department and then were retained in the Phycher archives as meta-data.
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25°C
- Humidity (%): 30-70%
- Air changes (per hr): at least ten changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (07.00 to 19.00) and twelve hours darkness.

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distilled

Details on oral exposure:

VEHICLE
- Concentration in vehicle:
In the first step of the study, 2.0003 g of the test item were weighed and distilled water was added to a 10 mL volumetric flask respectively. The preparation was stirred by vortex to obtain a blue solution just before the administration.
In the second step of the study, 0.3005 g of the test item were weighed and distilled water was added to a 10 mL volumetric flask respectively. The preparation was stirred by vortex to obtain a blue solution just before the administration.
In the third and fourth steps of the study, 0.0501 g and 0.0506 g of the test item were weighed and distilled water was added to two 10 mL volumetric flasks respectively. The preparations were stirred by vortex to obtain blue solutions just before the administration.
Each preparation was administered under a volume of 10 mL/kg body weight using a suitable syringe graduated fitted with an oesophageal metal canula.
- Amount of vehicle (if gavage): 10 mL/kg body weight
- Justification for choice of vehicle: most suitable formulation at the requested concentrations.

Doses:

2000, 300 and 50 mg/kg bw

No. of animals per sex per dose:

3 (6 for 50 mg/kg bw dose)

Control animals:

yes

Remarks:

The study identified No. TAO423-2017-005 was performed to assess the comportment of the strain of rat used at this laboratory in this environment and to give additional historical data.

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Systematic examinations were carried out to identify any behavioural or toxic effects on the major physiological functions 30 min, 1h, 3h, 4h, 24h, 48h after administration of the test item and continued daily during 14 days.
The animals were weighed on day D0 (just before administering the test item) then on day 2, day 7, and day 14.
- Necropsy of survivors performed: yes.
On day 14, the animals were euthanized sodium pentobarbital (Dolethal®). Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.
- Other examinations performed: clinical signs, body weight

Sex:

female

Dose descriptor:

LD50

Effect level:

> 50 - < 300 mg/kg bw

Based on:

test mat.

Mortality:

Two mortalities were noted in animals treated at the dose of 2000 mg/kg body weight at 3 hours post- dose.
Two mortalities were noted in animals treated at the dose of 300 mg/kg body weight, one at 3 hours post-dose and the second at 22 hours and 34 minutes post-dose.
No mortality was noted in animals treated at the dose of 50 mg/kg body weight

Clinical signs:

other: At 2000 mg/kg bw, the mortalities were preceded by an absence of spontaneous activity (2/2), muscle tones (2/2), Preyer’s reflex (2/2), righting reflex (2/2) associated with dyspnoea (2/2), eyes partly closed (2/2) and an increase of salivation (2/2). In

Gross pathology:

At 2000 mg/kg bw, rigor mortis were noted before the necropsy (2/2). The macroscopic examination of the animals revealed a thinning of forestomach (2/2) and blue coloration of stomach and intestine (2/2). The macroscopic examination of the surviving animal at the end of the study revealed a thinning of forestomach.
At 300 mg/kg bw, rigor mortis were noted before the necropsy (1/2). The macroscopic examination of the animals revealed a thinning of forestomach (1/2), blue coloration of stomach and intestine (1/2), blue corpus (1/2), blue liquid in stomach (1/2) and dark red spots on liver (1/2). The macroscopic examination of the surviving animal at the end of the study did not reveal treatment related changes.
At 50 mg/kg bw, the macroscopic examination of these animals at the end of the study did not reveal treatment related changes.

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The LD50 of the test item Reaction product of copper sulfate and triethylenetetramine is higher than 50 mg/kg body weight and lower than 300 mg/kg by oral route in the rat.
In accordance with the O.E.C.D. Test Guideline No. 423, the LD50 cut-off of the test item may be considered as 300 mg/kg body weight by oral route in the rat.
According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with the Regulation EC No. 1272/2008, the test item Reaction product of copper sulfate and triethylenetetramine has to be classified in category 3. The signal word “Danger” and hazard statement H301 “Toxic if swallowed” are required.

Executive summary:

The test item Reaction product of copper sulfate and triethylenetetramine was administered to a group of 3 female Sprague Dawley rats at the dose of 2000 mg/kg body weight, then to a group of 3 female Sprague Dawley rats at the dose of 300 mg/kg body weight and then to a group of 6 female Sprague Dawley rats at the dose of 50 mg/kg body weight. The experimental protocol was established according to the official method as defined in the O.E.C.D. Test Guideline No. 423 dated December 17th, 2001 and the test method B.1tris of the Council regulation No. 440/2008.

Two mortalities were noted in animals treated at the dose of 2000 mg/kg body weight at 3 hours post- dose.
The mortalities were preceded by an absence of spontaneous activity (2/2), muscle tones (2/2), Preyer’s reflex (2/2), righting reflex (2/2) associated with dyspnoea (2/2), eyes partly closed (2/2) and an increase of salivation (2/2).

Rigor mortis were noted before the necropsy (2/2).
The macroscopic examination of the animals revealed a thinning of forestomach (2/2) and blue coloration of stomach and intestine (2/2).
In the surviving animals (1/3), an absence of spontaneous activity, muscle tones, Preyer’s reflex, and righting reflex associated with dyspnoea, eyes partly closed and piloerection. The animal recovered a normal activity at 48 hours post-dose.
The macroscopic examination of this animal at the end of the study revealed a thinning of forestomach.

Two mortalities were noted in animals treated at the dose of 300 mg/kg body weight, one at 3 hours post-dose and the second at 22 hours and 34 minutes post-dose.
The mortalities were preceded by a decrease or an absence of spontaneous activity (2/2), muscle tones (2/2), Preyer’s reflex (2/2), righting reflex (2/2) associated with eyes partly closed (1/2) and piloerection (1/2).

Rigor mortis were noted before the necropsy (1/2).
The macroscopic examination of the animals revealed a thinning of forestomach (1/2), blue coloration of stomach and intestine (1/2), blue corpus (1/2), blue liquid in stomach (1/2) and dark red spots on liver (1/2).
In the surviving animals (1/3), an absence of spontaneous activity, muscle tones, Preyer’s reflex, and righting reflex associated with eyes partly closed and piloerection. The animal recovered a normal activity at 24 hours post-dose.
The macroscopic examination of this animal at the end of the study did not reveal treatment related changes.

No mortality was noted in animals treated at the dose of 50 mg/kg body weight
No clinical signs related to the administration of the test item were observed during the study.
The body weight evolution of the animals treated at the dose of 50 mg/kg body weight remained normal during the study.
The macroscopic examination of these animals at the end of the study did not reveal treatment related changes.

In conclusion, the LD50of the test item Reaction product of copper sulfate and triethylenetetramine is higher than 50 mg/kg body weight and lower than 300 mg/kg by oral route in the rat.
In accordance with the O.E.C.D. Test Guideline No. 423, the LD50cut-off of the test item may be considered as 300 mg/kg body weight by oral route in the rat.

According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with the Regulation EC No. 1272/2008, the test item Reaction product of copper sulfate and triethylenetetramine has to be classified in category 3. The signal word “Danger” and hazard statement H301 “Toxic if swallowed” are required.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw