Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-12-13 to 2013-01-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study reliable without restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: solid
Details on test material:
- Name of test material (as cited in study report): C40B.10-PTL Cobalt Borate Propionate
- Physical state: Solid blue/purple
- Stability: test substance was expected to be stable for the duration of testing.
- Storage condition of test material: the test substance was stored at room temperature.
- Solubility: soluble in hydrocarbons

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories, Inc.
- Age at study initiation: 8 - 11 weeks
- Weight at study initiation: 171 - 211 grams
- Fasting period before study: prior to each dosing, experimentally naive rats were fasted overnight. Feed was replaced approximately 3-4 hours after dosing.
- Housing: the animals were singly housed in suspended stainless steal caging with mesh floors, which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals (Natl. Res. Council, 2011).
- Diet (ad libitum, except during fasting): Harlan Teklad Global 16% Protein Rodent Diet® #2016
- Water (ad libitum): filtered tap water
- Acclimation period: 6 -27 days

Enrichment (e.g. toy) was placed in each cage.
during the fasting period, the rats were examined for health and weighed (initial).

ENVIRONMENTAL CONDITIONS
- Temperature: 19-23°C
- Relative humidity: 19-49%; the humidity was below the targeted lower limit for 3 days during the study. A portable humidifier was used to raise the humidity levels during this time.
- Air changes: 14/per hour
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
DOSAGE PREPARATION:
Prior to use, the test substance was ground in a coffee mill (Cuisinart, Model #DCG-20N). The ground sample was administered as a 40% w/w mixture in distilled water. A tissue homogenizer (Tissue tearor, Biospec, model 985370) was used to facilitate the preparation ofa homogeneous mixture. Preliminary solubility testing conducted by the laboratory, indicated that mixtures in excess of 40 (i.e., 45 - 80%) were too viscous to be administered properly.

MAXIMUM DOSE VOLUME APPLIED:
Individual doses were calculated based on the initial body weights, taking into account the density (1.003 g/mL; determined by the laboratory) and concentration of the test mixture.

PROCEDURE:
Initially, a single animal received a limit dose of 5,000 mg/kg. Due to mortality of this animal, a Main Test was conducted. For the Main Test, the test substance was administered in sequence to the animals (please refer to "Any other information on materials and methods incl. tables"). The decision to proceed with the next animal was based on the survival of the previous animal following dosing. Dose progressions and stopping criteria were determined using the statistical program The Acute Oral Toxicity (Guideline 425) Statistical Program (Westat, version 1.0, May 2001).
Doses:
55, 175, 550, 1750 and 5000 mg/kg
No. of animals per sex per dose:
9 female rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: individual body weights of the animals were recorded prior to test substance administration (initial) and again on Days 7 and 14 (termination) following dosing or after death. The animals were observed for mortality, sings of gross toxicity, and behavioural changes during the first several hours post-dosing and at least once daily thereafter for 14 days after dosing or until death occurred.
- Necropsy of survivors performed: yes
Surviving rats were euthanized via CO2 inhalation at the end of the 14-day observation period. Gross necropsies were performed on all decedents and euthanized animals. Tissues and organs of the thoracic and abdominal cavities were examined.
Statistics:
The Acute Oral Toxicity (Guideline 425) Statistical Program (Westat, version 1.0, May 2001) was used for all data analyses including: dose progression selections, stopping criteria determinations and/or LD50 and confidence limit calculations.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
310.2 mg/kg bw
Based on:
test mat.
Remarks on result:
other: LD50 based on maximum likelihood, & on assumed sigma of 0.5. Approx. 95% profile-likelihood based confidence interval of 0 mg/kg to > 20000 mg/kg & an approx. 90% profile-likelihood based confidence interval of 38.02 mg/kg to > 2530 mg/kg.
Mortality:
- 55 mg/kg dose level (1 animal):
This animal survived test substance administration.
- 175 mg/kg dose level (3 animals):
One animal was found dead within 3 hours of test substance administration.
- 550 mg/kg dose level (3 animals):
Two animals were found dead within 2.5 hours of test substance administration.
- 1750 mg/kg dose level (1 animal)
This animal was found dead within 40 minutes following test substance administration.
- 5000 mg/kg dose level (1 animal)
This animal was found dead within one hour of test substance administration.
Clinical signs:
- 55 mg/kg dose level (1 animal):
This animal appeared active and healthy during the study. There were no signs of gross toxicity, adverse pharmacologic effects, or abnormal behaviour.
- 175 mg/kg dose level (3 animals):
Clinical signs of the deceased animal prior to death included prone posture and irregular respiration. The clinical sign observed for one of the surviving animals following administration was soft faeces, but the animal recovered from this symptom by Day 2 and appeared active and healthy for the remainder of the 14-day observation period.
- 550 mg/kg dose level (3 animals):
Prior to death, the clinical signs noted for the two animals included irregular respiration, prone posture and/or hypoactivity. Following administration, the clinical signs for the surviving animal included reduced faecal volume, nasal discharge, ocular discharge, diarrhoea, hypoactivity, hunched posture and ano-genital staining. However, the animal recovered from these symptoms by Day 8, and appeared active and healthy for the remainder of the 14-day observation period.
- 1750 mg/kg dose level (1 animal)
There were no clinical signs noted for this animal prior to death.
- 5000 mg/kg dose level (1 animal)
There were no clinical sings noted for this animal prior to death.
Body weight:
- 55 mg/kg dose level (1 animal):
This animal gained body weight.
- 175 mg/kg dose level (3 animals):
The two surviving animals gained body weight.
- 550 mg/kg dose level (3 animals):
The surviving animal lost weight by day 7. It showed a continued weight gain thereafter through Day 14.
Gross pathology:
- 55 mg/kg dose level (1 animal):
No gross abnormalities were noted for this animal when necropsied at the conclusion of the 14-day observation period.
- 175 mg/kg dose level (3 animals):
Gross necropsy of the decedent revealed red lungs. No gross abnormalities were noted for the euthanized animals when necorpsied at the conclusion of the 14-day observation period.
- 550 mg/kg dose level (3 animals):
Gross necropsy of the decedents revealed distention of the stomach. No gross abnormalities were noted for the euthanized animal when necropsied at the conclusion of the 14-day observation period.
- 1750 mg/kg dose level (1 animal)
Gross necropsy of the decedent revealed distention of the stomach.
- 5000 mg/kg dose level (1 animal)
Gross necropsy of the decedent revealed distention of the stomach.

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
LD50 (female rats): 310.2 mg/kg bw
According to the EC-Commission directive 67/548/EEC and its subsequent amendments, the test substance is classified as harmful if swallowed.
According to the EC-Regulation 1272/2008 and subsequent regulations, the test item is classified as Category 4.