Reactive Blue F08-0170

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 October 2010 - 20 October 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to EU, OECD & US EPA test standards in compliance with GLP and reported with a valid GLP certificate.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Details on test material:

Substance Name: Reactive Blue F08-0170

Test animals

Species:

rat

Strain:

other: Wistar RjHan:WI

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test Animals

Species and strain: RjHan:(WI) Wistar rats
Source: Laboratoire Elevage Janvier, B.P. 4105, Route des Chênes Secs, 53940 Le Genest-St-Isle CEDEX FRANCE
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies
Number of animals: 5 animals/sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at study start: Young adult rats
Body weight range at dosing: Between 210 g and 250 g
Acclimatization time: 6 days
 
Husbandry

Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
Room-Box: 242/1
Housing: Individual caging
Cage type: Type II. polypropylene/polycarbonate
Bedding: Laboratory bedding: Lignocel Hygienic Animal Bedding produced by J. Rettenmaier & Söhne GmbH+Co.KG (Holzmühle 1, 73494 Rosenberger, Germany); A copy of the relevant Certificate of Analysis is maintained in LAB Research Ltd.'s archive.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20.4- 24.9 °C
Relative humidity: 31 - 62 %
Ventilation: 15-20 air exchanges/hour
Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.

The temperature and relative humidity was recorded twice daily during the study.


Food and Water Supply

Animals received ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottle ad libitum. The food is not considered to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at LAB Research Ltd.

Identification

The individual identification was performed using numbers written on the tail with a marker pen. The numbers were given on the basis of LAB Research Ltd.' s Master File for each animal allocated to the treatment groups. The cages were identified by cards containing information about study code, sex, dose group, cage number and individual animal numbers.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment. The test item was applied as a single dose as supplied to the shaved skin and remained in contact with the skin for the 24- hour exposure period. For that purpose, the appropriate amount of the test item was moistened with distilled water and distributed as uniformly as possible. Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin by a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.

At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.

Duration of exposure:

24 hours

Doses:

The test item was not expected to be lethal at 2000 mg/kg bw. A limit test was therefore performed.

No. of animals per sex per dose:

5 males/5 females per dose

Control animals:

no

Details on study design:

Clinical Observations

Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the 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.


Measurement of Body Weight

The body weights were recorded on Day 0 (before test item administration) and on Days 3, 7 and 14.


Necropsy

All animals were anaesthetised with Euthasol 40% and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.

Statistics:

None

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No effects.

Mortality:

No mortality occurred after a 24-hour dermal exposure of REACTIVE BLUE F08-0170 administered at 2000 mg/kg bw to RjHan:(WI) Wistar rats followed by a 14-day observation period.

Clinical signs:

other: No clinical signs were observed after the treatment with the test item or during the 14 day observation period.

Gross pathology:

The single oral gavage of Reactive Blue F08-0170 at a dose level of 2000 mg/kg bw, was not associated with any test item-related gross findings.

Other findings:

Bluish staining was recorded on the skin in all animals after dosing. The discoloration of the skin lasted 1 day in male and up to 7 days in female rats. No other local dermal signs were observed.

Any other information on results incl. tables

INDIVIDUAL CLINICAL OBSERVATIONS STUDY CODE: 10/252-002P TEST SYSTEM: Rj Han: WI Rat TEST ITEM: Reactive Blue F08-0170

Dose Level: 2000mg/kg

Sex: Male

  

Cage No.	Animal No.	Observations	Observation days																Frequency
			0		1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	2189	Symptom Free	+	+	-	+	+	+	+	+	+	+	+	+	+	+	+	+	15/16
		Skin (Bluish)	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	1/16
2	2190	Symptom Free	+	+	-	+	+	+	+	+	+	+	+	+	+	+	+	+	15/16
		Skin (Bluish)	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	1/16
3	2191	Symptom Free	+	+	-	+	+	+	+	+	+	+	+	+	+	+	+	+	15/16
		Skin (Bluish)	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	1/16
4	2192	Symptom Free	+	+	-	+	+	+	+	+	+	+	+	+	+	+	+	+	15/16
		Skin (Bluish)	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	1/16
5	2193	Symptom Free	+	+	-	+	+	+	+	+	+	+	+	+	+	+	+	+	15/16
		Skin (Bluish)	-	-	+	-	-	-	-	-	-	-	-	-	-	-	-	-	1/16

 

Dose Level: 2000mg/kg

Sex: Female

Cage No.	Animal No.	Observations	Observation days																Frequency
			0		1	2	3	4	5	6	7	8	9	10	11	12	13	14
6	2194	Symptom Free	+	+	-	-	-	-	+	+	+	+	+	+	+	+	+	+	12/16
		Skin (Bluish)	-	-	+	+	+	+	-	-	-	-	-	-	-	-	-	-	4/16
7	2195	Symptom Free	+	+	-	-	-	-	+	+	+	+	+	+	+	+	+	+	12/16
		Skin (Bluish)	-	-	+	+	+	+	-	-	-	-	-	-	-	-	-	-	4/16
8	2196	Symptom Free	+	+	-	-	-	-	-	-	-	+	+	+	+	+	+	+	9/16
		Skin (Bluish)	-	-	+	+	+	+	+	+	+	-	-	-	-	-	-	-	7/16
9	2197	Symptom Free	+	+	-	-	-	-	-	-	-	+	+	+	+	+	+	+	9/16
		Skin (Bluish)	-	-	+	+	+	+	+	+	+	-	-	-	-	-	-	-	7/16
10	2198	Symptom Free	+	+	-	-	-	+	+	+	+	+	+	+	+	+	+	+	13/16
		Skin (Bluish)	-	-	+	+	+	-	-	-	-	-	-	-	-	-	-	-	3/16

Remarks:       +: present                                                                           absent

h=hour (s)                                                                 Treatment day= Day 0

Frequency of observation = number of occurence of observation / total number of observations

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute dermal median lethal dose (LD50) of the test item REACTIVE BLUE F08-0170 was found to be higher than 2000 mg/kg body weight in male and female RjHan:(WI) Wistar rats. The substance is not classified.

Executive summary:

An acute dermal toxicity study was performed with test item REACTIVE BLUE F08-in RjHan:(WI) Wistar rats, in compliance with OECD Guideline No.: 402.

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied, moistened with distilled water, as a single dermal 24-hour exposure followed by a 14‑day observation period.

Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 3, 7 and 14. Rats were euthanized and a gross macroscopic examination performed at the end of the 2-week observation period (Day 14).

The results of the study were summarized as follows:

 

Mortality

 

No mortality occurred.

 

Systemic clinical signs

 

No clinical signs were observed after the treatment with the test item or during the 14‑day observation period.

 

Local dermal signs

 

No local dermal signs were observed during the entire study period. However, bluish staining was observed on the skin in all animals after dosing from Day 1 to Day 7.

 

Body weight

 

The body weight and body weight gain of REACTIVE BLUE F08-0170 treated animals did not show any test item-related effect.

 

Necropsy

 

There was no evidence of Reactive Blue F08-0170-related findings at necropsy. Uterus in estrus was observed in one female 2196.

The substance is not classified as harmful by dermal exposure.