Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Study conducted between 25th February 2014 and 20th March 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (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:
Identification: FAT 20341/A TE
Physical State/Appearance: black powder, solid at 20 °C
Batch: BOP 02-12 (Navy PLK 241, BS)
Purity: 80.1%
Expiry date: 21 November 2017
Storage Conditions: room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Female Wistar (RccHan™:WIST) strain rats were supplied by Harlan Laboratories UK Ltd., Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non-pregnant. After an acclimatization period of at least five days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The body weight variation did not exceed ± 20% of the body weight of the initially dosed animal.

The animals were housed in groups of up to three in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

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.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Test item preparation:
For the purpose of the study the test item was freshly prepared, as required, as a solution in distilled water.

The test item was formulated within two hours of being applied to the test system. It is assumed that the formulation was stable for this duration.

No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.

Procedure:
Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose.

Groups of fasted animals were treated as follows:

Dose Level (mg/kg) Concentration (mg/mL) Dose Volume (mL/kg) Number of Rats
2000 200 10 3
2000 200 10 3

Dosing:
All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each group to confirm the survival of the previously dosed animals.

Doses:
2000 mg/kg
No. of animals per sex per dose:
3 animals per dose group (all females)
Control animals:
no
Details on study design:
- Duration of observation period following administration:
The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for up to fourteen days.

- Frequency of observations and weighing:
Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment or at death.

- Necropsy of survivors performed: yes

- Other examinations performed: clinical signs, body weight, necropsy.

Rats are the preferred species of choice as historically used for safety evaluation studies and are specified in the appropriate test guidelines.

Results and discussion

Preliminary study:
Not applicable.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 - < 5 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal was found dead one day after dosing. Please see table 1 for more information in the any other information on results section.
Clinical signs:
Clinical Observations. Signs of systemic toxicity noted in the animal that died during the study were hunched posture, lethargy, pilo erection and cyanosis. Diarrhea was noted in the second group of three animals. Blue stained urine and feces was noted in all animals. All visible skin and eyes of four animals were stained blue.
Body weight:
Body Weight. Surviving animals showed expected gains in body weight over the observation period, except for one animal which showed expected gain in body weight during the first week but body weight loss during the second week.
Gross pathology:
Necropsy findings:
Discoloration of the heart, lungs, liver, spleen, kidneys, gastric mucosa, non glandular epithelium of the stomach, small and large intestines and bladder and dark blue colored liquid present in the stomach were noted at necropsy of the animal that died during the study. Discoloration of the heart, liver, kidneys, gastric mucosa, non glandular epithelium of the stomach and small and large intestines were noted at necropsy of one animal that was killed at the end of the study. Discolored kidneys were noted at necropsy of two animals that were killed at the end of the study. No abnormalities were noted at necropsy of two animals that were killed at the end of the study. Please see table 2 in the any other information on results section for more details.

Any other information on results incl. tables

TABLES

Table 1     Individual Clinical Observations and Mortality Data

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Female

0

0UF

0UF

0UF

0UF

0F

0

0

0

0

0

0

0

0

0

0

0

0

1-1

Female

0

0UF

0UF

0UF

0UF

0F

0

0

0

0

0

0

0

0

0

0

0

0

1-2

Female

HL

HLPCyUF

HLCyUF

HPLUF#

X

 

 

 

 

 

 

 

 

 

 

 

 

 

2-0

Female

0

0

DUF

DUF

0UF#

0F#

0F#

0F#

0F#

0F#

0F#

0#

0#

0#

0#

0#

0#

0#

2-1

Female

0

0

DUF

DUF

0UF#

0F#

0F#

0F#

0F#

0F#

0F#

0

0

0

0

0

0

0

2-2

Female

0

0

DUF

DUF

DUF#

0F#

0F#

0F#

0F#

0F#

0F#

0#

0#

0#

0#

0#

0#

0#



0= No signs of systemic toxicity      H = Hunched posture                        L = Lethargy        P = Pilo‑erection                 Cy = Cyanosis           D = Diarrhea

F= Faeces stained dark blue             U =Blue stained urine                       # = Skin and eyes stained blue                        X = Animal dead

Table 2     Individual Necropsy Findings:

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

1-0 Female

Killed Day 14

No abnormalities detected

1-1 Female

Killed Day 14

No abnormalities detected

1-2 Female

Found dead Day 1

Heart: discolored

Lungs: discolored

Liver: discolored

Spleen: discolored

Kidneys: discolored

Stomach: dark blue colored liquid present

Gastric mucosa: discolored

Non-glandular epithelium of the stomach: discolored

Small intestine: discolored

Large intestine: discolored

Bladder: discolored

2-0 Female

Killed Day 14

Heart: discolored

Liver: discolored

Kidneys: discolored

Gastric mucosa: discolored

Non-glandular epithelium of the stomach: discolored

Small intestine: discolored

Large intestine: discolored

2-1 Female

Killed Day 14

Kidneys: discolored

2-2 Female

Killed Day 14

Kidneys: discolored

 

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System - Category 5, >2000 – 5000 mg/kg body weight).

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

Executive summary:

Introduction:

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

 

Methods:

A group of three fasted females was treated with the test item at a dose level of 2000 mg/kg body weight. This was followed by a further group of three fasted females at the same dose level. Dosing was performed sequentially.

 

The test item was administered orally as asolutionindistilled water. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

Results:

Mortality. One animal was found dead one day after dosing.

 

Clinical Observations. Signs of systemic toxicity noted in the animal that died during the study were hunched posture, lethargy, pilo‑erection and cyanosis. Diarrhea was noted in the second group of three animals. Blue stained urine and feces was noted in all animals. All visible skin and eyes of four animals were stained blue.

 

Body Weight. Surviving animals showed expected gains in body weight over the observation period, except for one animal which showed expected gain in body weight during the first week but body weight loss during the second week.

 

Necropsy. Discoloration of the heart, lungs, liver, spleen, kidneys, gastric mucosa, non‑glandular epithelium of the stomach, small and large intestines and bladder and dark blue colored liquid present in the stomach were noted at necropsy of the animal that died during the study. Discoloration of the heart, liver, kidneys, gastric mucosa, non‑glandular epithelium of the stomach and small and large intestines were noted at necropsy of one animal that was killed at the end of the study. Discolored kidneys were noted at necropsy of two animals that were killed at the end of the study. No abnormalities were noted at necropsy of two animals that were killed at the end of the study.

Conclusion:

The acute oral median lethal dose (LD50) of teh test item in the female Wistar strain rat was astimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System - Category 5, >2000 - 5000 mg/kg body weight).

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.