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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 February 2012 to 7 March 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in accordance with OECD and EU Guidance with GLP certificate. Rated as Klimisch 2 because it is a read-across study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Product name: Bismuth Subsalicylate

Test animals

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.

Administration / exposure

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

Results and discussion

Preliminary study:
Not applicable
Effect levels
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

Applicant's summary and conclusion

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.