Reactive Blue F08-0170

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

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:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class 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: RjHan:WI

Sex:

female

Details on test animals or test system and environmental conditions:

Test Animals:

Species and strain: RjHan:WI rats
Source: Laboratoire Elevage Janvier, B.P. 4105, Route des Chênes Secs, 53940 Le Genest-St-Isle CEDEX FRANCE
Hygienic level at supplier: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals, 3 animals/group
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, 9-10 weeks old
Date of receipt: 23 September 2010
Body weight at treatment: 211 – 223 g
Acclimation period: At least 12 days

Husbandry

Animal health: Only healthy animals were used for the test. The veterinarian certified the health status.
Number of animal room: 522/7
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: Lignocel Bedding for Laboratory Animals was available to animals during the study.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3°C
Relative humidity: 30 - 70%
Ventilation: 15-20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

The temperature and relative humidity were 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 considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

The supplier provided an analytical certificate for the batch used. 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.

Animal Identification

Animals were individually identified using numbers written on the tail with an indelible 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, with information about study code, sex, dose group, cage number and individual animal numbers.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

distillied

Details on oral exposure:

Justification of the dose:

The initial dose level was selected on the basis of the information provided by the Sponsor. The LD50 value was expected to be above 2000 mg/kg bw.

Initially, three female animals were treated with 2000 mg/kg bw of Reactive Blue F08 0170. As no mortality occurred within 24 hours after dosing, a second group of three animals received 2000 mg/kg bw Reactive Blue F08 0170 approximately 24 hours after treatment of the first group. No mortality occurred in the second treatment group. Hence, further testing was not required according to the test guidelines. The test was terminated on completion of the 14-day observation period.

Doses:

Single dose, 2000 mg/kg bw

No. of animals per sex per dose:

3 initial; 3 additional.

Control animals:

no

Details on study design:

Procedure

A single oral dose was administered by gavage followed by a fourteen-day observation period. The animals were fasted for about 16 hours prior to treatment. The food but not the water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment. A constant treatment volume of 10 mL/kg bodyweight was applied.

Clinical Observations

Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. 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.


Body Weight Measurement

The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (day 0) and weekly thereafter.


NECROPSY

Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthasol® 40 %; details are presented in 3.1.3.). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

EVALUATION OF THE RESULTS


The method used was not intended to allow the calculation of a precise LD50 value.

The test item was ranked into categories of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423.

Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.

Statistics:

None.

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: No effects noted.

Mortality:

Reactive Blue F08-0170 did not cause mortality at a dose level of 2000 mg/kg bw.

Clinical signs:

other: Treatment with Reactive Blue F08-0170 caused decreased activity (3/6) 2 to 4 hours after treatment. No clinical signs were observed during the 14 days observation period. Bluish urine and faeces were observed in the bedding of Group 1; bluish faeces and p

Gross pathology:

A single oral gavage of Reactive Blue F08-0170 to the RjHan: WI rat at a dose level of 2000 mg/kg followed by a 14-day observation period did not result in any test item related macroscopic findings.

Other findings:

None

Any other information on results incl. tables

CLINICAL OBSERVATIONS

DOSE LEVEL: 2000 mg/kg									SEX: FEMALE
	Animal Number		Observation days
Cage No.		Observations	0						1-14	Frequency
			30' 1h 2h 3h 4h 6h
	1826	Symptom Free	+	+	-	-	+	+	+	18/20
		Activity decreased	-	-	-1	-1	-	-	-	2/20
1*	1827	Symptom Free	+	+	-	-	+	+	+	18/20
		Activity decreased	-	-	-1	-1	-	-	-	2/20
	1828	Symptom Free	+	+	-	+	+	+	+	19/20
		Activity decreased	-	-	-1	-	-	-	-	1/20
DOSE LEVEL: 2000 mg/kg									SEX: FEMALE
	Animal Number		Observation days
Cage No.		Observations	0						1-14	Frequency
			30' 1h 2h 3h 4h 6h
	1829	Symptom Free	+	+	+	+	+	+	+	20/20
2**	1830	Symptom Free	+	+	+	+	+	+	+	20/20
	1831	Symptom Free	+	+	+	+	+	+	+	20/20
Remarks:		+: present	-: absent
		h=hours (s)	Treatment day= Day 0
		Frequency of observation = number of occurence of observation / total number of
		observations
		*: Urine and faeces coloured (bluish) in litter 3h Day 0
		**: Urine and faeces coloured (bluish) in litter 3h Day 0
		Urine coloured (purple) in litter 6h Day 0

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 item Reactive Blue F08-0170 was found to be above 2000 mg/kg bw in female RjHan: WI rats. The substance is not classified.

Executive summary:

The single-dose oral toxicity of Reactive Blue F08-0170 was performed according to the acute toxic class method (OECD 423, OPPTS 870.1100 and Commission Regulation (EC) No 440/2008, B.1 tris (L 142, 30 May 2008)) in RjHan: WI rats.

 

Two groups of three female RjHan: WI rats (9-10 weeks of age) were treated with Reactive Blue F08-0170 at a dose level of 2000 mg/kg bw (Group 1 and Group 2).

 

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. As no mortality occurred in this dose group within 24 hours after dosing, a confirmatory treatment was performed on 3 further females (Group 2) at the same dose level (2000 mg/kg bw). Rats were maintained without compound administration for a 2‑week observation period after the day of dosing. As no mortality was observed in the second dose group, no further treatment was needed.

 

Treatment was carried out by gavage for each animal after an overnight food withdrawal (about 16 hours prior to treatment). Food was made available again 3 hours after the treatment.Reactive Blue F08-0170 was administered as a solution prepared in distilled water at a concentration of 200 mg/ml at a dosing volume of 10 ml/kg bw.

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.

 

Results

 

Mortality

 

Reactive Blue F08-0170 did not cause mortality at a dose level of 2000 mg/kg bw.

 

Clinical observations

 

Treatment with Reactive Blue F08-0170 caused decreased activity (3/6) 2 to 4 hours after treatment. No clinical signs were observed during the 14 days observation period. Bluish urine and faeces were observed in the bedding of Group 1; bluish faeces and purple urine were noted in the bedding of Group 2 on Day 0.

 

Body weight and body weight gain

 

Body weight gains of Reactive Blue F08-0170-treated animals showed no indication of a treatment-related effect during the entire study period.

The substance is not classified as harmful by oral exposure.