Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-10-27 till 2009-11-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study under GLP without deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

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
Details on test material:
- Name of test material (as cited in study report): FAT 40849/A TE
- Substance type: reactive dyestuff for cellulose fibers
- Physical state: blue powder
- Analytical purity: 76.9% of all colored components
- Lot/batch No.: TZ 5463 / BOP 03-09
- Expiration date of the lot/batch: 2014-06-30
- Storage condition of test material: At room temperature at about 20 °C

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Harlan Laboratories B.V., Kreuzelweg 53, 5961 NM Horst / The Netherlands
- Age at study initiation: 10 weeks
- Weight at study initiation: 164.4 - 197.2g
- Fasting period before study:overnight prior to intubation (ca 18-19h)
- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding
- Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 30/09 (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) ad libitum (except for the overnight fasting period prior to intubation and approximately 3-4 hours post dose).
- Water: Community tap water from Füllinsdorf ad libitum.
- Acclimation period: Group 1: Oct 27 to Nov 3, Group 2 Oct 27 to Nov 5

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): relative humidity between 30-70% (values above 70% during cleaning process possible)
- Air changes (per hr): Air-conditioned with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): automatically controlled light
cycle of 12 hours light and 12 hours dark, music
during the daytime light period.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.2 g/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: vehicle chosen after non-GLP solubility trial before the study initiation date
- Purity: Purified water prepared at Harlan Laboratories Ltd. was used as vehicle (deionised water which was processed and treated by the PURELAB Option-R unit, which links four purification technologies: reverse osmosis, adsorption, ion-exchange and photo oxidation).

MAXIMUM DOSE VOLUME APPLIED:
The animals received a single dose of the test item by oral gavage administration at 2000 mg/kg body weight after being fasted for approximately 18 to 19 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing.
The dosing volume was 10 mL/kg body weight.


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: experience with simular substances
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
6 females in two groups of three
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Viability / Mortality:
Daily during the acclimatization period, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (in common with the clinical signs) and twice daily during days 2-15.

Clinical Signs:
Daily during the acclimatization period, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1, depending on the occurrence of clinical signs of toxicity. Once daily during days 2-15.

Body Weights:
On test days 1 (prior to administration), 8 and 15.

- Necropsy of survivors performed: yes. All animals were killed at the end of the observation period by carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained.
Statistics:
no statistical analysis was performed

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: CL not applicable
Mortality:
No intercurrent deaths occurred during the course of the study.
Clinical signs:
Slight sedation and slightly ruffled fur were observed in all animals within the first 30 minutes after treatment and the slightly ruffled fur persisted up to the 5-hour observation (Group 2) or all animals at the 3- and 5-hour readings. Black to dark blue faeces were noted at day 2 up to day 3 or 4.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
Pathology, Macroscopic Findings: Black brown discolouration of the kidneys was recorded in all animals at necropsy.

Any other information on results incl. tables

Body Weight (g)

Dose mg/kg

Animal No.

Sex

Day 1 (treatment)

Day 8

Day 15

2000

1

F

164.4

188.1

192.2

2

F

197.2

215.6

221.0

3

F

186.8

198.3

214.1

2000

4

F

182.9

203.9

216.6

5

F

188.8

215.3

219.7

6

F

179.5

194.7

211.0

Macroscopial findings

Animal

Organ

Finding

Necropsy

1

Kidneys

Discoloration, black brown

Nov 18

2

Kidneys

Discoloration, black brown

Nov 18

3

Kidneys

Discoloration, black brown

Nov 18

4

Kidneys

Discoloration, black brown

Nov20

5

Kidneys

Discoloration, black brown

Nov20

6

Kidneys

Discoloration, black brown

Nov20

Mortality 

Dose

Ani-

 

 

 

 

 

 

mg/kg

mal

Sex

Death Date

Day

P

K

S

bw

No.

 

 

 

 

 

 

2000

1

F

18-Nov-2009

15

X

 

 

 

2

F

18-Nov-2009

15

X

 

 

 

3

F

18-Nov-2009

15

X

 

 

2000

4

F

20-Nov-2009

15

X

 

 

 

5

F

20-Nov-2009

15

X

 

 

 

6

F

20-Nov-2009

15

X

 

 

Key:  P planned necropsy; K killedin extremis, S spontaneous death

Clinical Signs

Dose

Ani-

 

 

Test days

 

 

 

 

 

 

 

 

 

 

 

 

 

 

mg/kg

mal

Sex

Signs

1

 

 

 

 

2

3

4

5

6

7

8

9

10

11

12

13

14

15

bw

No.

 

 

0.5*

1*

2*

3*

5*

 

 

 

 

 

 

 

 

 

 

 

 

 

 

2000

1

F

No clinical signs

 

 

 

 

 

 

 

 

X

X

X

X

X

X

X

X

X

X

X

 

 

 

Ruffled fur

1

1

1

1

1

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sedation

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Dark red eyes

 

 

 

X

X

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Black to dark blue feces

 

 

 

 

 

X

X

X

 

 

 

 

 

 

 

 

 

 

 

 

2

F

No clinical signs

 

 

 

 

 

 

 

 

X

X

X

X

X

X

X

X

X

X

X

 

 

 

Ruffled fur

1

1

1

1

1

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sedation

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Dark red eyes

 

 

 

X

X

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Black to dark blue feces

 

 

 

 

 

X

X

X

 

 

 

 

 

 

 

 

 

 

 

 

3

F

No clinical signs

 

 

 

 

 

 

 

 

X

X

X

X

X

X

X

X

X

X

X

 

 

 

Ruffled fur

1

1

1

1

1

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sedation

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Dark red eyes

 

 

 

X

X

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Black to dark blue feces

 

 

 

 

 

X

X

X

 

 

 

 

 

 

 

 

 

 

 

Key:       1 slight (max. grade of 3), X noted

*             Examinations were performedwithinthe first 30 minutes and at approximately 1, 2, 3 and 5 hours after treatment.

No clinical signs were evident in any animal during the acclimatization period.


Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The median lethal dose of FAT 40849/A TE after single oral administration to female rats, observed over a period of 14 days, is:
LD50 (female rat): greater than 2000 mg/kg body weight
Based upon the classification criteria according to „Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008“ the test item is not classified in respect to the acute oral toxicity study in rats.
Executive summary:

Two groups, each consisting of three female RccHan:WIST (SPF) rats, were treated with FAT 40849/A TE by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was formulated in purified water at a concentration of 0.2 g/mL and administered at a dosing volume of 10 mL/kg.

 

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded within the first 30 minutes and approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

 

No intercurrent deaths occurred during the course of the study.

 

Slight sedation and slightly ruffled fur were observed in all animals within the first 30 minutes after treatment and the slightly ruffled fur persisted up to the 5-hour observation (Group 2) or day 2 (Group 1). Dark red eyes were noted in all animals at the 3- and 5-hour readings. Black to dark blue faeces were noted at day 2 up to day 3 or 4.

 

The body weight of the animals was within the range commonly recorded for this strain and age.

 

Black brown discolouration of the kidneys was recorded in all animals at necropsy.

 

The median lethal dose of FAT 40849/A TE after single oral administration to female rats, observed over a period of 14 days, is:

LD50 (female rat): greater than 2000 mg/kg body weight

 

Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, the test item is not classified in respect to the acute oral toxicity study in rats.