Tetracopper hexahydroxide sulphate

Endpoint summary

Description of key information

Acute toxicity: Oral

A study was performed to assess the acute oral toxicity of the test material according to OECD 423. Animals treated with 2000 mg/kg were found dead during the day of dosing or one day after dosing. No deaths were noted in animals treated with 200 mg/kg. Signs of systemic toxicity noted in females were hunched posture, pilo-erection and diarrhoea.

The acute median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl:CD® (SD) IGS BR) strain rat was estimated to be in the range of 300 - 500 mg/kg bodyweight.

 

Acute toxicity: Dermal

A study was performed to assess the acute dermal toxicity of the test material according to OECD 423.

There were no deaths on the study and no clinical observations or signs dermal irritation. The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl: CD® (SD) IGS BR) strain rat was found to be greater than 2000 mg/kg bodyweight.

 

Acute toxicity: Inhalation

Three groups of rats, consisting of five male and five female rats per group were used for the main study. Rats from groups I, II, and III were exposed to breathing zone concentration levels 3.038, 2.069 and 1.194 mg test item/L of air, respectively. Rats from all groups were exposed for 4 h and surviving rats were observed for a period of 14 days.

Sign of toxicity lethargy was observed in rats exposed with the test item.

Percent mortalities observed (both sexes combined) were 90, 30, and 10 at the breathing zone concentration levels 3.038, 2.069, and 1.194 mg/L air of test item, respectively which indicates dose dependent mortality.

Group I: A decrease in the body weight, was observed in surviving male rat on days 1 and 3 which increased on days 7 and 14 when compared with day 0 body weight.

Group II:A decrease in the mean body weight, was observed in surviving rats on days 1, 3 and 7 which increased on day 14 when compared with day 0 mean body weight in both the sexes.

Group III: A decrease in the mean body weight, was observed on days 1 and 3 which increased on days 7 and 14 when compared with day 0 body weight in male rats. While, in case of female rats a decrease in the mean body weight, was observed on days 1, 3 and 7 which increased on day 14 when compared with day 0 mean body weight.

External examination of the found dead and terminally sacrificed rats did not reveal any abnormality of pathological significance. Visceral examination of the found dead rats revealed liver: reddish discolouration (Rat N º 1, 3, 4 and 9) and lungs: multiple whitish foci (Rat N º 13, 17, 19 and 24) whereas other found dead and terminally sacrificed rats did not reveal any lesion of pathological significance.

Lesions observed in the found dead rats could be correlated with the test item used in the present study.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental phase: 07 February 2002 to 28 February 2002. Report issue: 12 June 2002.

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

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:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Male and female Sprague-Dawley CD (Crl: CD® (SD) IGS BR) strain rats were supplied by Charles River (UK) Ltd, Margate, Kent, UK.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Approximately eight weeks old
- Weight at study initiation: At least 200g
- Fasting period before study: Overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: The animals were housed in groups of three by sex in solid-floor polypropylene cages furnished with woodflakes
- Diet: provided ad libitum (except for immediately prior to and after dosing)
- Water: provided ad libitum (except for immediately prior to and after dosing)
- Acclimation period: At least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Air changes (per hr): At least 15
- Photoperiod (hrs dark / hrs light): Twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
- Environmental enrichment: The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5%

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/ml for 2000 mg/kg dose and 200 mg/ml for 200 mg/kg dose.
- Amount of vehicle (if gavage): 10 ml/kg

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

Doses:

2000 mg/kg and 200 mg/kg

No. of animals per sex per dose:

2000 mg/kg: 3 female
200 mg/kg: 3 female and 3 male

Control animals:

no

Statistics:

None

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 300 - < 500 mg/kg bw

Based on:

test mat.

Mortality:

Animals treated with 2000 mg/kg were found dead during the day of dosing or one day after dosing. No deaths were noted in animals treated with 200 mg/kg.

Clinical signs:

other: Signs of systemic toxicity noted in females were hunched posture, pilo-erection and diarrhoea. Additional signs of systemic toxicity noted in animals treated at a dose level of 2000 mg/kg were lethargy, decreased respiratory rate, laboured respiration and

Gross pathology:

Abnormalities noted at necropsy of animals that died during the study were haemorrhagic or abnormally red lungs, dark liver, dark kidneys, blue liquid present in the stomach, epithelial sloughing of the gastric mucosa and non-glandular region of the stomach and haemorrhagic small intestine. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl:CD® (SD) IGS BR) strain rat was estimated to be in the range of 300 - 500 mg/kg bodyweight.

Executive summary:

Introduction

The study was performed to assess the acute oral toxicity of the test material. The method used was designed to meet the requirements of the following guidelines:

• OECD Guidelines for the Testing of Chemicals No. 423 "Acute Oral Toxicity - Acute ToxicClass Method" (adopted 22 March 1996)

• Commission Directive 96/54/EC Method B1 tris Acute Oral Toxicity (Oral - Acute ToxicClass Method)

Method

A group of three fasted females was treated with the test material at a dose level of 2000 mg/kg bodyweight. Based on the results from this dose level further groups of fasted animals were treated at a dose level of 200 mg/kg bodyweight. Dosing was performed sequentially. The test material was administered orally as a dispersion in 0.5% carboxy methyl cellulose.

Results

Animals treated with 2000 mg/kg were found dead during the day of dosing or one day after dosing. No deaths were noted in animals treated with 200 mg/kg.

Signs of systemic toxicity noted in females were hunched posture, pilo-erection and diarrhoea. Additional signs of systemic toxicity noted in animals treated at a dose level of 2000 mg/kg were lethargy, decreased respiratory rate, laboured respiration and ataxia. One female treated with 2000 mg/kg was found comatose four hours after dosing. All females treated with 200 mg/kg recovered one day after dosing. No signs of systemic toxicity were noted in males treated with 200 mg/kg.

Bodyweight. The surviving animals showed expected gains in bodyweight.

Conclusion

The acute median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl:CD® (SD) IGS BR) strain rat was estimated to be in the range of 300 - 500 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

traditional method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Species: Rat (Rattus norvegicus)
Strain: Wistar
Age/Weight at Dosing: 10 to 12 weeks
Weight (g): Male: Minimum: 292.2, Maximum: 359.4 ; Female: Minimum: 184.9, Maximum: 225.1
Source: Animal Breeding Facility, Jai Research Foundation
Total Number of: Thirty (15 males and 15 females)
Animals Used Female rats were nulliparous and non-pregnant
The study was undertaken in compliance with the guidelines of the “Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC), International” and “Guidelines for Laboratory Animals Facility” issued by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India.

ENVIRONMENTAL CONDITIONS
Animal Room: BMR 29, Department of Toxicology
Temperature: 19 to 23 °C
Relative Humidity: 56 to 66%
Air Changes: Minimum 15 air changes/hour
Photoperiod: The photoperiod was 12 hours artificial light and 12 hours darkness, light hours being 06:00 h – 18:00 h which was maintained through automatic timer.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

>= 1 - <= 4 µm

Details on inhalation exposure:

- Experimental Design
The 4 hour acute inhalation exposure was carried out using nose-only inhalation exposure equipment supplied by CH Technologies, USA. This dynamic inhalation exposure system consists of a flow-past nose-only inhalation exposure chamber with Perspex rat exposure/restraint tubes, air compressor, flow meter, rotating brush generator (dry powder), cascade impactor, temperature and humidity probe (thermo-hygrometer), oxygen monitor, carbon dioxide monitor, and open face gravimetric filter sampler.

- Inhalation Equipment
The rats were exposed using a nose-only inhalation exposure system with a dynamic air flow rate of 727 to 728 air changes per hour, ensuring an adequate oxygen content of at least 19% and a homogeneous, evenly distributed, respirable test aerosol with a mass median aerodynamic diameter (MMAD) particle size ranged between 1-4 microns. An exposure atmosphere CO2 level was less than 1%.

- Description of Inhalation Chamber
The dynamic inhalation equipment has 2 main parts namely outer plenum and inner plenum. Inner plenum volume is 862 cm3 and outer plenum volume is 1,200 cm3 so the total volume = 2062 cm3 = 2.062 Liters. The exposure unit (outer plenum) is made of stainless steel with 12 portholes to accommodate transparent perspex rat exposure tubes. These exposure tubes are accommodated in the portholes of the inhalation chamber. The adjustable unit of the exposure tube is set in such a way that rats breathe the test item aerosol through the window panel of the exposure tube. Observations during the inhalation experiment are made through transparent perspex rat exposure tubes. The outlet unit connected to a suction pump. The outgoing air from the chamber passes through an impinger containing 1.0% sodium hydroxide solutions and moisture traps.

- Description of Dust Generator System
The dust generator system is manufactured by PALAS GmbH and supplied by CH Technologies (USA), Inc. The dust generator system is a long assembly of tubing having several tube fittings attached. The feed system consists of a feed stock reservoir, generating system consists of a brush housing with integrated brush, spacer holder, up/down schalter, inlet of air for diserpsion, connection of compressed air with reducer, switch for manual or external controls (air, brush speed, schalter speed).

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

The nominal concentration of aerosolized substance was calculated to be 10.277, 6.646 and 3.446 mg/L air for group I, II and III, respectively, based on the mass of test item which used during exposure and the total chamber air flow during exposure.

No. of animals per sex per dose:

In the main test, 5 males and 5 females were tested per dose.

Control animals:

no

Details on study design:

OBSERVATIONS
- Toxic Signs
All rats were observed for any signs of toxicity and mortality hourly, during the 4 h exposure period and at 1 and 2 h after the exposure on the day of exposure. Subsequently, surviving rats were observed twice a day for morbidity and mortality for a period of 14 days following exposure. Clinical signs were recorded once a day.
- Body Weight
Body weights were recorded prior to exposure on day 0 and on days 1, 3, 7, and 14 and at death.
- Necropsy
At the end of the study, all survived rats were sacrificed by intraperitoneal administration of thiopentone sodium and subjected to gross pathological examination. All found dead rats were also subjected to gross pathological examination. This consisted of an external examination and the opening of the nasal passage, abdominal, and thoracic cavities. The appearance of any macroscopic abnormality was recorded.

Statistics:

As this study was conducted as a full study at three different breathing zone concentrations, the acute inhalation median lethal concentration (LC50) of the test item was calculated from the observed mortality data by the Probit analysis (Finney, 1971) method.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

ca. 2.091 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

ca. 2.314 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

Percent mortalities observed (both sexes combined) were 90, 30, and 10 at the breathing zone concentrations 3.038, 2.069, and 1.194 mg/L air of the test item, respectively which indicates dose dependent mortality (see table1).

Clinical signs:

other:

Body weight:

Group I: A decrease in the body weight, was observed in surviving male rat on days 1 and 3 which increased on days 7 and 14 when compared with day 0 body weight.
Group II: A decrease in the mean body weight, was observed in surviving rats on days 1, 3 and 7 which increased on day 14 when compared with day 0 mean body weight in both the sexes.
Group III: A decrease in the mean body weight, was observed on days 1 and 3 which increased on days 7 and 14 when compared with day 0 body weight in male rats. In case of female rats a decrease in the mean body weight, was observed on days 1, 3 and 7 which increased on day 14 when compared with day 0 mean body weight.

Gross pathology:

Necropsy (Macroscopic Findings):
- External: External examination of the found dead and terminally sacrificed rats did not reveal any abnormality of pathological significance.
- Internal: Visceral examination of the found dead rats revealed liver: reddish discolouration (Rat Nº 1, 3, 4 and 9) and lungs: multiple whitish foci (Rat Nº 13, 17, 19 and 24) whereas other found dead and terminally sacrificed rats did not reveal any lesion of pathological significance. Lesions observed in the found dead rats could be correlated with the test item used in the present study.

Other findings:

External examination of the found dead and terminally sacrificed rats did not reveal any abnormality of pathological significance. Visceral examination of the found dead rats revealed liver: reddish discolouration (Rat N º 1, 3, 4 and 9) and lungs: multiple whitish foci (Rat N º 13, 17, 19 and 24) whereas other found dead and terminally sacrificed rats did not reveal any lesion of pathological significance.

Interpretation of results :
The calculated acute inhalation median lethal concentration (LC50) value of the test item for combined sex was 2.091 mg/L air with 95% lower fiducial limit of 1.717 mg/L air and upper fiducial limit of 2.548 mg/L air. The regression equation is y = -15.264 + 6.103 x.
The calculated acute inhalation median lethal concentration (LC50) value of the test item for male rats was 2.314 mg/L air with 95% lower fiducial limit of 1.524 mg/L air and upper fiducial limit of 3.515 mg/L air. The regression equation is y = -8.001 + 3.864 x.

Text Table: 1 Concentration, Mortality/Rats Treated

Breathing ZoneConcentration (mg/L air)	Males (mortality/total)	Females (mortality/total)
3.038	4/5	5/5
2.069	1/5	2/5
1.194	1/5	0/5

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The calculated acute inhalation median lethal concentration (LC50) value of test item for combined sex was 2.091 mg/L air with 95% lower fiducial limit of 1.717 mg/L air and upper fiducial limit of 2.548 mg/L air. The regression equation is y = -15.264 + 6.103 x.
The calculated acute inhalation median lethal concentration (LC50) value of the test item for male rats was 2.314 mg/L air with 95% lower fiducial limit of 1.524 mg/L air and upper fiducial limit of 3.515 mg/L air. The regression equation is y = -8.001 + 3.864 x.

Executive summary:

Threegroups of rats, consisting of five male and five female rats per group were used for the main study. Rats from groups I, II, and III were exposed to breathing zone concentration levels 3.038, 2.069 and 1.194 mg test item/L of air, respectively. Rats from all groups were exposed for 4 h and surviving rats were observed for a period of 14 days.

Sign of toxicity lethargy was observed in rats exposed with the test item.

Percent mortalities observed (both sexes combined) were 90, 30, and 10 at the breathing zone concentration levels 3.038, 2.069, and 1.194 mg/L air of test item, respectively which indicates dose dependent mortality.

Group I: A decrease in the body weight, was observed in surviving male rat on days 1 and 3 which increased on days 7 and 14 when compared with day 0 body weight.

Group II:A decrease in the mean body weight, was observed in surviving rats on days 1, 3 and 7 which increased on day 14 when compared with day 0 mean body weight in both the sexes.

Group III: A decrease in the mean body weight, was observed on days 1 and 3 which increased on days 7 and 14 when compared with day 0 body weight in male rats. While, in case of female rats a decrease in the mean body weight, was observed on days 1, 3 and 7 which increased on day 14 when compared with day 0 mean body weight.

External examination of the found dead and terminally sacrificed rats did not reveal any abnormality of pathological significance. Visceral examination of the found dead rats revealed liver: reddish discolouration (Rat N º 1, 3, 4 and 9) and lungs: multiple whitish foci (Rat N º 13, 17, 19 and 24) whereas other found dead and terminally sacrificed rats did not reveal any lesion of pathological significance.

Lesions observed in the found dead rats could be correlated with the test item used in the present study.

Breathing Zone Concentration (mg/L Air)	N° of Animals Used	Mortalities/Sex		Mortalities %
		Male	Female
3.038	5 Males + 5 Females	04	05	90
2.069	5 Males + 5 Females	01	02	30
1.194	5 Males + 5 Females	01	00	10

The calculated acute inhalation median lethal concentration (LC₅₀) value of the test item for combined sex was 2.091 mg/L air with 95% lower fiducial limit of 1.717 mg/L air and upper fiducial limit of 2.548 mg/L air. The regression equation is y = -15.264 + 6.103 x.

The calculated acute inhalation median lethal concentration (LC₅₀) value of test item for male rats was 2.314 mg/L air with 95% lower fiducial limit of 1.524 mg/L air and upper fiducial limit of 3.515 mg/L air. The regression equation is y = -8.001 + 3.864 x.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

2 091 mg/m³ air

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:

Experimental phase:11 February 2002 to 25 February 2002. Report issue: 12 June 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Male and female Sprague-Dawley CD (Crl: CD® (SD) IGS BR) strain rats were supplied by Charles River (UK) Ltd, Margate, Kent, UK.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Approximately eight weeks old
- Weight at study initiation: At least 200g
- Fasting period before study: None
- Housing: The animals were housed in groups of five by sex in solid-floor polypropylene cages furnished with woodflakes except during the 24-hour period when they were housed individually.
- Diet: provided ad libitum
- Water: provided ad libitum
- Acclimation period: At least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Air changes (per hr): At least 15
- Photoperiod (hrs dark / hrs light): Twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
- Environmental enrichment: The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Type of coverage:

semiocclusive

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5% used to moisten the test material

Details on dermal exposure:

TEST SITE
- % coverage: Approximately 10% of the total body surface ara.
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage.
- Application site: Area of shorn skin

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Cotton wool moistened with distilled water was used to remove any residual test material.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 200 mg/kg
-- For solids, paste formed: test material was moistened with 0.5% carboxy methyl cellulose (CMC)

Duration of exposure:

24-hours

Doses:

2000 mg/kg bodywieght

No. of animals per sex per dose:

5 male and 5 female

Control animals:

no

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no unscheduled deaths on the study.

Clinical signs:

other: There were no signs of systemic toxicity

Gross pathology:

No gross pathological abnormalities were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl: CD® (SD) IGS BR) strain rat was found to be greater than 2000 mg/kg bodyweight.

Executive summary:

Introduction

The study was performed to assess the acute dermal toxicity of the test material in the Sprague-Dawley CD (Crl: CD® (SD) IGS BR) strain rat. The method was designed to meet the requirements of the following guidelines:

 

• OECD Guidelines for the Testing of Chemicals No. 402 "Acute Dermal Toxicity" (adopted 24 February 1987)

 

• Commission Directive 92/69/EEC Method B3 Acute Toxicity (Dermal)

 

 

Method

A group of ten animals (five males and five females) was given a single, 24-hour, semi-occluded dermal application of test material to intact skin at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

 

Results

There were no deaths on the study and no clinical observations or signs of dermal irritation.

No abnormalities were noted at necropsy.

All animals showed expected gains in bodyweight over the study period.

 

Conclusion

The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl: CD® (SD) IGS BR) strain rat was found to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Justification for classification or non-classification

The acute median lethal dose (LD50) of the test material in the Sprague-Dawley CD (Crl:CD® (SD) IGS BR) strain rat was estimated to be in the range of 300 - 500 mg/kg bodyweight which according to Table 3.1.2 in Regulation (EC) No 1272/2008 corresponds to a classification of Acute Oral Category 4 (H302). This classification is in line with the harmonised classification published in Annex VI of the CLP Regulation.