Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
The study will be conducted according to the following guideline:
Acute Oral Toxicity-Acute Toxic Class Method, OECD Guideline for the Testing of Chemicals No 423, Adopted: 17th December 2001
Guidance Document on Acute Oral Toxicity Testing, OECD Series on Testing and Assessment No 24, 2001
Guidance Document on the Recognition, Assessment and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation. Environmental Health and Safety Publications Series on Testing and Assessment No 19, 2000
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
Details on test material:
1 MATERIALS AND METHODS
1.2 Test Item
Designation in Test Facility: 17031402G
Date of Receipt: 14. Mar. 2017
Condition at Receipt Room temperature, in proper conditions
1.2.1 Specification.
Name Blendazol Red Blendwell
Batch no. E 328
Appearance Dark Red Powder
Composition 2-Naphtalenesulfonic acid, 7-amino-4-hydroxy-3,8-bis[[4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]-, triso dium salt
Purity 95 % by HPLC
Homogeneity homogeneous
Expiry date 17. Feb. 2019
Storage Room Temperature: (20 ± 5°C); Keep away from humidity
CAS No. 607724-47-0
EINECS-No. 612-028-6
Chemical Class not stated
Stability H2O: 96h; EtOH: unknown; acetone: unknown CH3CN: unknown; DMSO: unknown
Solubility H2O: to be determined*; EtOH: unknown; acetone: unknown CH3CN: unknown; DMSO: unknown


* Will be determined in another GLP-study at LAUS GmbH and will be stated in the final report, this
This information is not provided by the sponsor but determined at LAUS GmbH.


Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Test Animals
8-12 weeks; 6 females Wistar rats from Dobrá Voda, Slovak Republic were used for test, they were non-pregnant and nulliparous
Health condition of animals was examined by a veterinarian before initiation of the study.
The animals were acclimated under the conditions identical to the conditions during the experiment 5 days prior to the start of treatment. The acclimation was according to the standard operation procedure.
The animals were housed in plastic cages suspended on stainless steel racks, up to 3 animals per cage in a room equipped with central air-conditioning. The average room temperature was maintained within the range of 22.9 ± 0.4° C, relative humidity within 53.8 ± 2 %. The light regimen was set to a 12-hour light /12-hour dark cycle. Sanitation was performed according to the standard operation procedures.
The laboratory food ssniff (Spezialdiäten GmbH, Germany) was offered at recommended doses each day approximately at the same time. The certificate of analysis is included in the raw data.
The animals received tap water for human consumption. Supply of drinking was unlimited. The quality of drinking water is periodical analysed and recorded; certificate of analysis is included in the raw data.
Bedding: Lignocel S3/4, Lufa - ITL GmbH, Germany
The animals in the cage were marked by a line (I-III) on the tail with a waterproof marker. Each cage was marked with the study code, ID of animals and date of administration of the test item.
Females were used for testing according to OECD TG 423 because females are typically the more sensitive gender.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Aqua pro injection, lot number 0125K16, Expiry date: 10/2019 Manufacturer: Imuna Pharm a.s., Slovakia Aqua pro injectione is a standard vehicle according to OECD TG 423
Details on oral exposure:
The required amount of the test item (according to the body weight and dose) was mixed with vehicle (aqua pro injectione) shortly before administration.
The test item was administered in a single dose by gavage using a metal stomach tube. Animals were fasted prior to dosing (food but not water were withheld over-night). Following a period of fasting, animals were weighed and the test item administered. After test item administration, food was withheld for further 3-4 hours
Doses:
The starting dose could be selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg body weight. Available information
indicated that the test item is likely to be non-toxic with regard to acute toxicity. A limit dose of 2000 mg/kg body weight was used as a starting dose.
No. of animals per sex per dose:
One group of 3 females was dosed. Test item-related mortality was not observed during 24 hours and therefore in a second step another 3 females were treated at the same dose
Control animals:
no
Details on study design:
Clinical Observation:
Animals were observed individually immediately after administration of the test item and 0.5, 1, 2, and 4 hours later. Each animal was inspected daily for the next 14 days.
Observations included: changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity, and behavioural pattern. Particular attention was given to potential neurologic endpoints such as tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body Weight:
Individual weights of animals were measured immediately prior to administration of the test item and weekly thereafter. Weight differences after first and second weeks after administration were calculated and recorded.
Necropsy
All test animals were subjected to gross necropsy and the results were recorded for each animal.

Statistics:
All (6/6 females) animals survived the limit dose of 2000 mg/kg body weight.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
>= 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No mortality was observed during the study. During the follow up period, no animals displayed signs of intoxication, change of health, nor any other adverse reaction
Clinical signs:
No observed signs
Body weight:
Stagnation of body weight in rats No 1, 2, 4, and 6 were observed between first and second week after administration.
Gross pathology:
All animals were necropsied. During necropsy, no macroscopic findings were observed.

Any other information on results incl. tables

Stagnation of body weight in rats No 1, 2, 4, and 6 were observed between first and second week after administration. Summary results of body weights are presented inTable3.

Table3 Body Weight

Sex

Dose

ID

Body Weight (g)

Body Weight Difference (g)

Initial

Week 1

Week 2

Week 1 - Initial

Week 2 - Initial

Week 2 - Week 1

2000 mg/kg

1

205

220

220

15

15

0

2

210

220

220

10

10

0

3

195

205

210

10

15

5

4

200

220

220

20

20

0

5

200

210

220

10

20

10

6

210

230

230

20

20

0

Table4Necropsy Results

Sex

Dose

ID

Result

Sex

Dose

ID

Result

2000 mg/kg

1

no finding

2000 mg/kg

4

no finding

2

no finding

5

no finding

3

no finding

6

no finding

1.1         Summary Results

The test item Blendazol Red Blendwellwas administeredto 6 females at a limit dose of 2000 mg/kg. Stagnation of body weight in 4 animals were observed between one and two weeks after administration of the test item.

Nosigns of toxicity were observed at the dosage of 2000 mg/kg during the first 4 hours in females or the 14-day observation period thereafter.During necropsy, no macroscopic findings were observed.

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The test item Blendazol Red Blendwell was administered to 6 females at a limit dose of 2000 mg/kg. Stagnation of body weight in 4 animals were observed between one and two weeks after administration of the test item.
No signs of toxicity were observed at the dosage of 2000 mg/kg during the first 4 hours in females or the 14-day observation period thereafter. During necropsy, no macroscopic findings were observed.
Executive summary:

The oral acuste toxicity of Blendazol Red Blendwell was determined following OECD Guideline 423 Acute Toxic Class Method after a single oral administration dose of 2000 mg/kg body weight in 6 femalles rats and observed for 14 days, obtaining that :

The LD50of the test item Blendazol Red Blendwell is greater than 2000 mg/kg body weight after single oral administration to Wistar rats.

Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423 it can be concluded that the test item Blendazol Red Blendwell is classified in Category 5/Unclassified with a LD50cutoff value equal to or greater than 5000 mg/kg body weight, after single oral administration to Wistar rats.