Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 February 2015 to 18 March 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

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
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA 712-C-98-190
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:
- Appearance: Violet powder
- Storage conditions: Controlled room temperature (15 to 25 °C, below 70 % relative humidity and protected from light and humidity)

Test animals

Species:
rat
Strain:
other: CRL:(WI)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: CRL:(WI) (outbred, SPF-quality)
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
- Age at study initiation: Approximately 10 weeks old
- Weight at study initiation: 178 to 232 g
- Fasting period: On the night before treatment, the animals were fasted
- Housing: 3 animals/cage in type II polypropylene/polycarbonate.
- Diet (e.g. ad libitum): Animals received ssniff, SM R/M "Autoclavable complete diet for rats and mice- breeding and maintenance” produced by ssniff , Spezialdiäten GmbH, D-59494 Soest Germany (Batch no.: 680 2237 expiry date: March 2015), ad libitum, and tap water from the municipal supply, as for human consumption from a 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.
- Acclimation period: 19/20 days


ENVIRONMENTAL CONDITIONS
- Temperature: 19.8 to 22.8 °C
- Humidity: 31 to 70 % (relative)
- Air changes: 15 – 20 air exchanges/hour
- Photoperiod: Lighting period; 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES:
- From: 12 February 2015
- To: 18 March 2015



Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle:
- Batch number: 2171213

CLASS METHOD
- Rationale for the selection of the starting 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.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 females; the test material was administered to two subsequent groups of three female rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observatins were made 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter.
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs, body weight, organ weights, histopathology, other: 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.
Statistics:
No statistical analysis was performed as the method used is not intended to allow the calculation of a precise LD50 value.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Acute oral administration of Acid Blue 249 did not caused any test item related toxic effect. Bluish discolouration of the faeces was observed on Day 0, Day 1 and Day 2.
Body weight:
Body weight gains of Acid Blue 249 treated animals during the study showed no indication of a test item-related effect.
Gross pathology:
There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

Any other information on results incl. tables

Table 1: Body Weight Data (g):         

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0

SEX: FEMALE

Cage No.

Animal Number

Body Weight (g)

Body Weight Gain (g)

Days

-1

0

7

14

 -1-0

0-7

7-14 

-1-14 

1

2211

233

217

246

250

-16

29

4

17

2212

242

232

250

245

-10

18

4

12

2213

244

228

253

255

-16

25

2

11

2

2214

237

229

257

258

-8

28

1

21

2215

229

219

243

247

-10

24

4

18

2216

187

178

207

215

-9

29

8

28

Mean:

228.7

217.2

242.7

246.5

-11.5

25.5

3.8

17.8

 

 

 

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 test material was found to be above 2000 mg/kg bw in female CRL:(WI) rats.
Executive summary:

The single-dose oral toxicity of the test material 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).

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 dose volume of 10 mL/kg bw.

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.

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.

The test material did not cause mortality at a dose level of 2000 mg/kg bw.

Acute oral administration of the test material did not cause any test item related toxic effect. Bluish discolouration of the faeces was observed on Day 0, Day 1 and Day 2.

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

There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

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

According to Annex I, Regulation (EC) No. 1272/2008, the test material is unclassified for acute oral exposure.