Reaction mass of dieuropium tricarbonate and digadolinium tricarbonate and disamarium tricarbonate

Endpoint summary

Description of key information

Acute Oral Toxicity

Under the conditions of this study, the acute oral LD50 value of the test material was > 2 000 mg/kg bw in rats.

Acute Inhalation Toxicity

Under the conditions of this study, the LC50 (4 hour) of the test material is in excess of 5.12 mg/L for male rats.

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:

12 September 2017 to 27 September 2017

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:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 11 weeks old
- Weight at study initiation: 230 - 243 g
- Fasting period before study: On the night before treatment, the animals were fasted. The food, but not water, was withheld during an overnight period. The food was returned 3 hours after the treatment.
- Housing: 3 animals/ cage in Type II polypropylene/polycarbonate cages
- Diet: Ad libitum, except for the night before treatment
- Water: Animals received tap water from the municipal supply from 500 mL bottles, ad libitum
- Acclimation period: At least 26 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 - 25.0 °C
- Humidity (%): 30 - 80 %
- Air changes (per hr): 15 - 20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m

Route of administration:

oral: gavage

Vehicle:

methylcellulose

Remarks:

1 % Methyl cellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test material was formulated in 1 % methyl cellulose at a concentration of 200 mg/mL

MAXIMUM DOSE VOLUME APPLIED: The dose volume was 10 mL/kg bw

DOSAGE PREPARATION: The test material was freshly formulated at a concentration of 200 mg/mL in the vehicle in the Pharmacy of Citoxlab Hungary Ltd. on the day of administration. The formulation container was stirred continuously with a magnetic stirrer until dose administration procedures were complete.

CLASS METHOD
- Rationale for the selection of the starting dose: The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. Limit dose of 2 000 mg/kg bw was selected as a starting dose. Initially, 3 female animals were treated at dose level of 2 000 mg/kg bw. No mortality was observed, therefore a further 3 animals were treated at the dose level of 2 000 mg/kg bw. As no mortality was observed in the second dose group, further testing was not required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).

Doses:

2 000 mg/kg bw

No. of animals per sex per dose:

6 animals, 3 animals/ group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.
- Necropsy of survivors performed: Yes. Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthanimal 40 %). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
- Other examinations performed: Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Statistics:

The method used was not intended to allow the calculation of a precise LD50 value.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed at dose level of 2 000 mg/kg bw.

Clinical signs:

All animals were symptom-free during the 14-day observation period at a dose level of 2 000 mg/kg bw.

Body weight:

There were no effects on body weights or body weight gains that could be attributed to treatment with the test material.

Gross pathology:

There was no evidence of the macroscopic observations in the animals dosed at 2 000 mg/kg bw and terminated on Day 14.

Interpretation of results:

other: Not classified in accordance with EU criteria.

Conclusions:

Under the conditions of this study, the acute oral LD50 value of the test material was > 2 000 mg/kg bw in rats.

Executive summary:

The single-dose oral toxicity of the test material was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in Crl:WI Wistar rats. Two groups of 3 female Crl:WI rats were treated with the test material at a dose level of 2 000 mg/kg bw (Group 1 and Group 2).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test material was formulated in 1 % methyl cellulose at a concentration of 200 mg/mL at a dose volume of 10 mL/kg bw.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy). All animals were subjected to a necropsy and a macroscopic examination.

Initially, three females (Group 1) were treated at a dose level of 2 000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required.

The results of the study were summarised as follows:

Mortality: No mortality was observed at dose level of 2 000 mg/kg bw.

Clinical Observations: All animals were symptom-free during the 14-day observation period at a dose level of 2 000 mg/kg bw.

Body Weight and Body Weight Gain: There were no effects on body weights or body weight gains that could be attributed to treatment with the test material.

Necropsy: There was no evidence of the macroscopic observations in the animals dosed at 2 000 mg/kg bw and terminated on Day 14.

Under the conditions of this study, the acute oral LD50 value of the test material was found to be above 2 000 mg/kg bw in female Crl:WI rats.

According to the GHS criteria, the test material can be ranked as "Unclassified" for acute oral exposure.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 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:

21 September 2018 to 23 October 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD Guidelines for Testing of Chemicals No. 433 “Acute Inhalation Toxicity: Fixed Concentration Procedure.”

Version / remarks:

Adopted 25 June 2018

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed concentration procedure

Limit test:

yes

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Test item prepared as follows:
Device used: Mechanical grinder
Number of bursts: Four
Burst time: 8 - 15 seconds
Method of sieving: Hand
Sieve used: 212 μm mesh

Species:

rat

Strain:

Wistar

Remarks:

RccHan™:WIST

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 76 to 82 days
- Weight at study initiation: 320 to 341 g
- Housing: The animals were housed five per cage. The cages were made of a polycarbonate body with a stainless steel mesh lid. Wood shavings (Lignocel 3/4) were used as bedding and were sterilised by autoclaving and changed at appropriate intervals each week. Cages, food hoppers and water bottles were changed at appropriate intervals.
- The cage of animals was provided with an Aspen chew block for environmental enrichment. Chew blocks were provided throughout the study and were replaced when necessary. The cage of animals was provided with a plastic shelter for environmental enrichment, which was replaced at the same time as the cage.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: The animals were allowed to acclimatize to the conditions described below for 12 days before treatment commenced.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 24 °C
- Humidity: 40 - 70 %
- Air changes: Each animal room was supplied with filtered fresh air, which was passed to atmosphere and not re-circulated.
- Photoperiod: Artificial lighting was controlled to give a cycle of 12 hours continuous light and 12 hours continuous dark per 24 hours.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

3.4 µm

Geometric standard deviation (GSD):

1.99

Remark on MMAD/GSD:

The mean MMAD value was within the ideal range of 1 to 4 microns, indicating the generated test material aerosol was respirable to the rat.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The exposure system consisted of a flow through nose only chamber and an aluminium alloy construction comprising a base unit, one animal exposure section with 20 exposure ports, and a top section. The dimensions of the chamber give an internal volume of 30 L. A curved aerosol conditioning pre-chamber was used on the exposure system, with an internal volume of 2.2 L. An elutriator, with an internal volume of 1.3 L, was connected to the aerosol generator. The elutriator connected to the pre-chamber by aerosol tubing.
- Exposure chamber volume: 33.6 L
- Method of holding animals in test chamber: Plastic nose-only restraint tube.
- Aerosol Generation: Solid Aerosol Generator: The SAG mechanism technique for the dispersion of dry dust and powders comprises two steps, the continuous supply of material to the disperser and the dispersal of the material as an aerosol. The method for metering powder to the disperser was to use a moving toothed belt. A regulated flow of compressed air conducted the aerosol to the inhalation chamber.
- Inlet Airflow: From in-house compressed air system (breathing quality). Flow: 28 L/minute
- Extract Airflow: Drawn by in-house vacuum system, filtered locally. Flow: 29 L/minute
- Temperature and humidity in air chamber: The mean chamber air temperature was 21.4 ± 0.35 °C and mean relative humidity was 52.2 ± 4.74 %.

TEST ATMOSPHERE
- Aerosol samples were collected as follows:
Sample type: Glass microfiber filter, held in an open face filter holder
Sample flow: 2.0 L/minute
Sample volume: Measured by wet-type gas metre
Sample frequency: 12 samples collected during exposure
Sample location: Animal exposure port
Sample analysis: Gravimetric
Aerosol samples were collected at irregular intervals due to additional samples being collected after adjustments made to operating conditions. A time-weighted average mean aerosol concentration was calculated.
- Particle Size Distribution was determined by cascade impaction – samples collected as follows:
Impactor type: Marple 290 Series Personal Cascade Impactor (Andersen Instruments, Smyrna, Georgia, USA)
Configuration: 298
Collection media: Stainless steel substrates and glass microfiber final stage filter
Sample flow: 2.0 L/minute
Sample volume: Measured by wet-type gas meter
Sample frequency: Two samples collected during exposure
Sample location: Animal exposure port
Sample analysis: Gravimetric, MMAD and GSD derived

DATA EVALUATION
- Determination of Time-Weighted Averages for Calculation of Aerosol Concentration of the test material: Samples of air were removed from the test chamber during exposure in order to determine the concentration of the test material in air. In the first instance, samples were obtained following equilibration and then at approximately 30-minute intervals. Additional samples were obtained as necessary to monitor the chamber concentration following adjustments to the exposure system. A time-weighted average was calculated from the individual data in order to prevent undue biasing of repeat samples in the overall mean.
Time-weighted average: sum of ‘weighted concentrations’ (sample concentrations weighted for time) calculated for each sampling occasion as follows:

‘Weighted concentration’ (mg/L) = (Concentration (mg/L) × ‘Time-weighting’ (min)) / Exposure duration (min)

Where the ‘time-weighting’ is the respective interval between adjustments to the exposure system or the midpoints between consecutive sampling occasions if no adjustment was made.

- Calculation of MMAD and GSD: MMAD and GSD - by linear regression of the probit of the cumulative percentage, by mass, of particles smaller than the ECD of each stage versus the logarithm of the ECD of each stage.

- Determination of Nominal Aerosol Calculation:
Nominal aerosol concentration (mg/L) = (Usage (g) × 1 000) / (Generation period (mins) × Airflow (L/minute))

- Determination of Chamber Air Changes:
Number of air changes per hour = (60 (minutes) × Airflow (L/minute)) / Chamber volume (L)

- Determination of T95 Equilibration Time:
Natural Logarithm (100 ÷ (100-95)) × ( Chamber volume (L) / Airflow (L/minute))

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Target: 5.0 mg/L
TWA Mean: 5.12 mg/L

No. of animals per sex per dose:

5 males

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Throughout the study, all cages were checked at least twice daily, once in the morning and again towards the end of the normal working day, for dead or moribund animals.
Clinical signs were recorded pre-exposure, at hourly intervals during exposure, immediately following exposure, 1 hour and 2 hours post-exposure and as late as possible in the working day.
During the observation period, the animals were observed once in the morning and once toward the end of the experimental day. On the day of study termination there was one observation (morning only).
Rats were weighed at least once during the week prior to exposure. The weight of each animal was recorded on Days 1 (prior to dosing), 2, 4, 8 and 15.
- Necropsy of survivors performed: Yes. At the end of the scheduled observation period, animals were killed by an overdose of pentobarbitone sodium followed by exsanguination.
All animals were subjected to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities. The macroscopic appearance of all examined organs was recorded. Tissues were discarded following necropsy.

Statistics:

In order to minimise the cumulative errors, which result from repeated rounding of numbers, some of the data in this report have been calculated continuously using unrounded numbers and only rounded for reporting. Consequently, any further calculation using these rounded numbers may include rounding errors in the last significant figure, possibly leading to small apparent discrepancies with other data in the report.

Days of pretreatment relate to study days, as follows:
Phase day P5
Day of study -8

Key result

Sex:

male

Dose descriptor:

LC50

Effect level:

> 5.12 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no unscheduled deaths.

Clinical signs:

other: There were no clinical signs related to treatment.

Body weight:

Slight body weight losses were evident for all animals on Day 2. Body weight gains were observed on Day 4 and continued to increase for the remainder of the observation period for all animals.

Gross pathology:

The macroscopic examination performed after a single exposure and a 14 day observation period revealed no test material related lesions. No abnormalities were seen in any of the animals.

Other findings:

Chamber Atmosphere Conditions
- The mean achieved concentration value was 102 % of the target concentration of 5 mg/L. The MMAD was within the ideal range of 1-4 microns for an acute inhalation study, indicating that the aerosol was respirable to the rat.

Interpretation of results:

other: Not classified in accordance with EU criteria

Conclusions:

Under the conditions of this study, the LC50 (4 hour) of the test material is in excess of 5.12 mg/L for male rats.

Executive summary:

The acute inhalation toxicity of the test material was investigated in accordance with the standardised guideline OECD 433, under GLP conditions.

One test group, consisting of five male RccHan™;WIST rats were exposed to the test material for a single 4-hour snout only exposure and observed for a period of fourteen days post exposure.

The mean achieved concentration value of the test material was 102 % of the target concentration of 5 mg/L. The MMAD was within the ideal range of 1-4 microns for an acute inhalation study, indicating that the aerosol was respirable to the rat.

There were no unscheduled deaths and no clinical signs related to treatment. Slight body weight losses were evident for all animals on Day 2. Body weight gains were observed on Day 4 and continued to increase for the remainder of the observation period for all animals. No macroscopic abnormalities were observed.

Under the conditions of this study, the LC50 (4 hour) of the test material is in > 5.12 mg/L for male rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute Oral Toxicity

The single-dose oral toxicity of the test material was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in Crl:WI Wistar rats. Two groups of 3 female Crl:WI rats were treated with the test material at a dose level of 2 000 mg/kg bw (Group 1 and Group 2).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test material was formulated in 1 % methyl cellulose at a concentration of 200 mg/mL at a dose volume of 10 mL/kg bw.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy). All animals were subjected to a necropsy and a macroscopic examination.

Initially, three females (Group 1) were treated at a dose level of 2 000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required.

The results of the study were summarised as follows:

Mortality: No mortality was observed at dose level of 2 000 mg/kg bw.

Clinical Observations: All animals were symptom-free during the 14-day observation period at a dose level of 2 000 mg/kg bw.

Body Weight and Body Weight Gain: There were no effects on body weights or body weight gains that could be attributed to treatment with the test material.

Necropsy: There was no evidence of the macroscopic observations in the animals dosed at 2 000 mg/kg bw and terminated on Day 14.

Under the conditions of this study, the acute oral LD50 value of the test material was found to be above 2 000 mg/kg bw in female Crl:WI rats.

According to the GHS criteria, the test material can be ranked as "Unclassified" for acute oral exposure.

Acute Inhalation Toxicity

The acute inhalation toxicity of the test material was investigated in accordance with the standardised guideline OECD 433, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

One test group, consisting of five male RccHan™;WIST rats were exposed to the test material for a single 4-hour snout only exposure and observed for a period of fourteen days post exposure.

The mean achieved concentration value of the test material was 102 % of the target concentration of 5 mg/L. The MMAD was within the ideal range of 1-4 microns for an acute inhalation study, indicating that the aerosol was respirable to the rat.

There were no unscheduled deaths and no clinical signs related to treatment. Slight body weight losses were evident for all animals on Day 2. Body weight gains were observed on Day 4 and continued to increase for the remainder of the observation period for all animals. No macroscopic abnormalities were observed.

Under the conditions of this study, the LC50 (4 hour) of the test material is > 5.12 mg/L for male rats.

Acute Dermal Toxicity

In accordance with Column 2 of REACH Annex VIII, information requirement section 8.5, in addition to the oral route, information shall be provided for at least one other route.  As data will be provided for the inhalation route, additional data via the dermal route is not required

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to acute toxicity via the oral or inhalation routes.