Registration Dossier

Administrative data

Description of key information

By read across to bismuth subsalicylate, bismuth trinitrate is not acutely toxic via the oral route. By read across to dibismuth trioxide, bismuth trinitrate is not acutely toxic via the inhalation route. Acute toxicity via the dermal route was not tested.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
21 February 2012 to 7 March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in accordance with OECD and EU Guidance with GLP certificate.
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: CRL:(WI)
Sex:
female
Details on test animals and environmental conditions:
EXPERIMENTAL ANIMALS
Species and strain: CRL:(WI) rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals, 3 animals/group
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, 8 weeks old
Body weight at treatment: 185 – 199 g
Acclimation period: At least 5 days

Husbandry

Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 522/8
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: Lignocel Bedding for Laboratory Animals was available to animals during the study. A copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m. Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

The temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply

Animals received ssniff® SM R/M-Z+H "Autoclavable complete diet for rats and mice
– breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Animal Identification

Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File, for each animal allocated to the treatment groups. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilles
Details on oral exposure:
Vehicle: Distilled water
Batch Number: 3450611
Expiry Date: 30 June 2014
Dose volume: 10 mL/kg bw

The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle, in the Central Dispensary Unit of CiToxLAB Hungary Ltd. on the day of administration. The formulation container was stirred continuously up to finishing the treatment.

Justification of the 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. In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose.

Initially, three female animals were treated with 2000 mg/kg bw of Bismuth Subsalicylate. No mortality was observed, therefore further 3 animals were treated at the dose level of 2000 mg/kg bw. As no mortality was observed in this 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:
2000 mg/kg bw
No. of animals per sex per dose:
6 females per dose
Control animals:
no
Details on study design:
Procedure

A single oral gavage administration was followed by a fourteen-day observation period. On the night before treatment, the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment.

OBSERVATIONS

Clinical Observations

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. 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.

Body Weight Measurement

The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly after.

NECROPSY

Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthasol® 40 %; Lot: 11H15 8; Expiry date: July 2014; Produced by: AST Beheer B.V. Oudewater Netherlands (Produlab Pharma, Raamsdonksveer)). 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.

EVALUATION OF THE RESULTS

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

The test item was ranked into categories of Globally Harmonized Classification
System (GHS) described in the OECD Guideline No. 423.

Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.
Statistics:
None
Preliminary study:
Not applicable
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Bismuth Subsalicylate did not cause mortality at a dose level of 2000 mg/kg bw.
Clinical signs:
Treatment with Bismuth Subsalicylate did not cause any clinical signs during the 14 days observation period.
Body weight:
Body weight gains of Bismuth Subsalicylate treated animals during the study showed no indication of a test item-related effect.
Gross pathology:
There was no evidence of the test item-related findings in animals dosed at 2000 mg/kg bw at necropsy.
Other findings:
None
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the read-across substance, Bismuth Subsalicylate, was found to be above 2000 mg/kg bw in female CRL:(WI) rats.
Executive summary:

The single-dose oral toxicity of Bismuth Subsalicylate 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) rats.

Two groups of three female CRL:(WI) rats were treated with the test item at a dose level of 2000 mg/kg bw (Group 1 and Group 2).

Initially, three females (Group 1) were treated at a dose level of 2000 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 according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.

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 item  was  administered  formulated  in  Distilled  water  at  a  concentration  of 200 mg/mL at a dosing 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 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

Results

Mortality

Bismuth Subsalicylate did not cause mortality at a dose level of 2000 mg/kg bw.

Clinical observations

Treatment with Bismuth Subsalicylate did not cause any clinical signs during the 14 days observation period.

Body weight and body weight gain

Body weight gains of Bismuth Subsalicylate treated animals during the study showed no indication of a test item-related effect.

Macroscopic Findings

There was no evidence of the test item-related findings in animals dosed at 2000 mg/kg bw at necropsy.

Conclusion:

Under the conditions of this study, the acute oral LD50 value of the test item Bismuth Subsalicylate was found to be above 2000 mg/kg bw in female CRL:(WI) rats.


Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
GLP study conducted according to OECD Guidance with a Klimisch score of 1.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
12 July 2010 to 26 July 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline study
Qualifier:
according to
Guideline:
OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
signed by Amt für Arbeitsschutz, Arbeitnehmerschutz Hamburg (2009-11-12)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: CD / Crl: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, D-97633 Sulzfeld
- Age at study initiation: 50 days (males), 64 days (females)
- Weight at study initiation: 229-242 g (males) and 236-240 g (females)
- Fasting period before study: Feeding was discontinued approx. 16 hours before exposure.
- Housing: During the 14-day observation period the animals are kept by sex in groups of 2 - 3 animals in MAKROLON cages (type III plus).
- Diet: Commercial diet, ssniff® R/M-H V1534 served as food.
- Water: ad libitum, tap water
- Acclimation period: At least 5 adaptation days and animals were acclimatised to the test apparatus for approx. 1 hour on 2 days prior to testing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 55 +/- 15
- Photoperiod: 12 hours dark/light cycle
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The study was carried out using a dynamic inhalation apparatus (air changes/h (≥ 12 times)) with a nose-only exposure of the animals according to KIMMERLE & EBEN. The apparatus consists of a cylindrical exposure chamber which holds the animals in pyrex tubes at the edge of the chamber in a radial position.
- Exposure chamber volume: 40 L
- Source and rate of air: Air was taken from the surrounding atmosphere of the laboratory room and filtered using an in-line disposable gas-filter. At the bottom of the exposure chamber, the air was sucked off at a lower flow rate than it was created by the dust generator in order to produce a homogenous distribution and a positive pressure in the exposure chamber (inflow 900 L/h, outflow 800 L/h, 22.5 air changes per hour).
- System of generating particulates/aerosols: The dust of the test material was generated with a rotating brush dust generator. The generator was fed with compressed air (5.0 bar) from a compressor.
- Treatment of exhaust air: The exhaust air was drawn through gas wash-bottles.
- Temperature, humidity, pressure in air chamber: Temperature (26.5°C ± 0.8°C) and humidity (69.2% ± 1.4%) were measured once every hour with a climate control monitor.

TEST ATMOSPHERE
- Brief description of analytical method used: The actual dust concentration in the inhalation chamber was measured gravimetrically with an air sample filter and pump, controlled by a rotameter. Dust samples were taken once every hour during the exposure.
- Samples taken from breathing zone: yes; a probe was placed close to the animals' noses and air was drawn through the air sample filter at a constant flow of air of 5 L/min for 1 minute. The filters were weighed before and after sampling (accuracy 0.1 mg).

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: An analysis of the particulate size distribution was carried out twice during the exposure period using a cascade impactor. The dust from the exposure chamber was drawn through the cascade impactor for 5 minutes at a constant flow rate of 5 L/min. The slides were removed from the impactor and weighed on an analytical balance. Delta of slides’ weight were determined.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The mass median aerodynamic diameter (MMAD) was estimated by means of non-linear regression analysis to be 2.700µm. The Geometric Standard Deviation (GSD) of the MMAD was calculated as 2.95 from the quotient of the 84.1%- and the 50%-mass fractions, both obtained from the above mentioned non-linear regression analysis. In addition, a sample of approx. 10 g test material was taken from the exposure chamber to determine the median physical particulate size with a Malvern Sizer by Malvern, 71083 Herrenberg, Germany.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: according to guideline
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Before initiating the study with the animals, a pre-test was carried out with the exposure system in order to verify that under the experimental settings chosen, the limit concentration of 5 mg/L air could be achieved by gravimetric analysis.
Duration of exposure:
4 h
Concentrations:
5.00 mg/L air (nominal concentration)
5.07 mg/L air (measured concentration)
No. of animals per sex per dose:
One concentration of 3 males and 3 females (limit test)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Careful clinical examinations were made at least twice daily until all symptoms subsided, thereafter each working day. Observations on mortality were made at least once daily (in the morning starting on test day 2) to minimize loss of animals to the study, e.g. necropsy or refrigeration of those animals found dead and isolation or sacrifice of weak or moribund animals. Individual weights of animals were determined once during the acclimatisation period, before and after the exposure on test day 1, on test days 3, 8 and 15. Changes in weight were calculated and recorded when survival exceeded one day. At the end of the test, all animals were weighed and sacrificed.
- Necropsy of survivors performed: yes
- Other examinations performed: Cageside observations included, but were not limited to: changes in the skin and fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, as well as somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremor, convulsions, salivation diarrhoea, lethargy, sleep and coma. The animals were also observed for possible indications of respiratory irritation such as dyspnoea, rhinitis etc.
Necropsy of all animals was carried out and all gross pathological changes were recorded. No microscopic examination was carried out as no pathological findings were noted at necropsy.
Statistics:
no data
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5.07 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No mortality occurred.
Clinical signs:
Under the present test conditions, a 4-hour inhalation exposure to dibismuth trioxide at a concentration of 5.07 mg/L air revealed slight ataxia and slight dyspnoea in 2 or 3 male and 3 female rats.
Body weight:
All animals gained the expected body weight throughout the study period.
Gross pathology:
No findings.
Other findings:
No other findings were observed.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the 4 -hour inhalation LC50 of dibismuth trioxide is >5.07 mg/L air, and hence, the LC50 cut-off value unclassified. According to the Globally Harmonized Classification System (GHS) the test item should be classified in the hazard category 'unclassified'.
Executive summary:

The aim of the present experiment was to obtain information on the acute toxicity and respiratory irritation, following a single 4-hour inhalation exposure of rats to dibismuth trioxide.

Rats were exposed to a dry aerosol of dibismuth trioxide at a gravimetricly determined concentration of 5.07±0.09 mg dibismuth trioxide/L air for 4 hours by inhalation using a dynamic nose-only exposure chamber. The aerosol was generated with the aid of a dry, rotating brush dust generator.

In the inhalation chamber, close to the animals' noses, the generated aerosol particulates had a mass median aerodynamic diameter (MMAD) of 2.700 µm as determined with a cascade impactor. The Geometric Standard Deviation (GSD) of the MMAD was calculated as 2.95. No smaller MMAD could be obtained with the test item supplied.

Under the present test conditions, a 4 -hour inhalation exposure to dibismuth trioxide at a concentration of 5.07 mg/L air (determined by gravimetric analysis) caused no mortality, and the body weight changes of all animals revealed throughout the study period normal gains.

A 4 -hour inhalation exposure to dibismuth trioxide at a concentration of 5.07 mg/L air revealed slight ataxia and slight dyspnoea on test day 1 immediately after end of exposure until 3 hours post exposure in 2 or 3 male and 3 female animals.

Under the conditions of this study, the 4 -hour inhalation LC50of dibismuth trioxide is >5.07 mg/L air, and hence, the LC50cut-off value is unclassified.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
5 070 mg/m³
Quality of whole database:
GLP study conducted according to OECD Guidance with a Klimisch score of 1.

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

Bismuth trinitrate is not acutely toxic via the oral route based on read-across to related substance, bismuth subsalicylate. In an acute oral toxicity study in rats, no deaths occurred at a bismuth subsalicylate dose of 2000 mg/kg bw. Therefore, the LD50 is considered to be > 2000 mg/kg bw.

In a supporting study, the acute oral toxicity in rats of read-across substance, bismuth was tested (Sano, 2005). No deaths occurred at a dose of 2000 mg/kg body weight. Therefore the LD50is considered to be >2000 mg/kg bw for both sexes. A read-across concept is applicable from bismuth to bismuth trinitrate considering that they both contain bismuth, the moiety of toxicological concern, and that the substances have similar physicochemical properties.

There are reports in the literature of human exposure to the related substance, bismuth(3+) tripotassium bis[2-hydroxypropane -1,2,3-tricarboxylate, by the oral route including case reports of poisoning incidents. It is difficult to draw any conclusions from these isolated case reports (see IUCLID Chapter 7.10).

In an acute inhalation toxicity test in rats with read-across substance, dibismuth trioxide (d50approximately 6 µm), no deaths occurred. The LC50was determined to be >5.07 mg/L air.

An acute dermal toxicity study with bismuth trinitrate is considered to be scientifically unjustified.


Justification for selection of acute toxicity – oral endpoint
Klimisch 1 study with read-across substance, bismuth subsalicylate

Justification for selection of acute toxicity – inhalation endpoint
Only 1 study available (for read-across substance, dibismuth trioxide).

Justification for selection of acute toxicity – dermal endpoint
An acute dermal toxicity study for bismuth trinitrate is considered unjustified since dermal uptake of bismuth is considered negligible (Annex VIII section 8.5 Column 2 of regulation (EC) 1907/2006).

Justification for classification or non-classification

Based on absence of adverse effects observed in an acute oral toxicity study with read-across substance, bismuth subsalicylate, and in an acute inhalation toxicity study with read-across substance, dibismuth trioxide, bismuth trinitrate is not classified for acute toxicity.