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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The aim of this study was to assess the toxicity potential of α,α-bis[4-(dimethylamino)phenyl]-4-(phenylamino)naphthalene-1-methanol (containing less than 0.1% Michler’s Ketone) Synonym Solvent Blue 4 (CAS No. – 6786-83-0) after single oral administration in rats and an observation period of 14 days.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):α,α-bis[4-(dimethylamino)phenyl]-4-(phenylamino)naphthalene-1-methanol (containing less than 0.1% Michler’s Ketone) Synonym Solvent Blue 4
- Substance type:Organic
-Purity:99.02%
- Storage condition of test material:Stored in cool, dry place. Kept in closed container when not in use.
- Other:
Handling and Disposal
Safety precautions: Avoided eye and skin contact. Avoided inhalation and spilling. Aprons, caps, mask, gloves and goggles were used to ensure the health and safety of the personnel.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation:8- 10 weeks at the time of dosing.
- Health Status :Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
- Weight at study initiation:Minimum: 131 g Maximum: 160 g (Individual body weights were within ± 7% prior to treatment after overnight fasting)
- Fasting period:were fasted for 16 to 18 hours, prior to dosing .
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding:All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) Batch No.: SPAR – 25/2014.
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No 400004..
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles
- Acclimation period:Animal nos. 1-3 were acclimatized for 5 days and 4-6 for 11 days prior to administration of the test item..

ENVIRONMENTAL CONDITIONS
-Temperature : Minimum: 19.80°C Maximum: 23.20°C
-Relative humidity:Minimum: 48.70 % Maximum: 69.20 %
-Light-dark-rhythm:12:12
-Air Changes : More than 12 changes per hour


Administration / exposure

Route of administration:
oral: unspecified
Vehicle:
other: distilled water
Details on oral exposure:
VEHICLE
- Amount of vehicle (if gavage):10 ml
- Justification for choice of vehicle:N/A
- Lot/batch no. (if required):N/A
- Purity:N/A

MAXIMUM DOSE VOLUME APPLIED:10 ml/kg body weight.

Doses:
G1 = 2000 mg/kg bw
No. of animals per sex per dose:
6 female rats
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
AAt the end of 14 day observation period, all surviving rats were euthanised by overdose of CO2 for external and internal observations.
- Other examinations performed:


Clinical Observation
After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all the six animals were observed once a day during the 14 day observation period.

Mortality
All the animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.

Body weight
All the rats were weighed on days 0 (prior to dosing), 7 and 14. Animals were weighed immediately after found dead.

Pathology
At the end of 14 day observation period, all surviving rats were euthanised by overdose of CO2 for external and internal observations

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed througout the experimentation period
Clinical signs:
At 2000mg/kg, animal no. 1 and 4 were observed with mild lethargy at 30 minutes, 1, 2, 3, 4 hours and on day 1 post dosing while additionally, animal no. 1 was observed with mild lethargy on day 2 and animal no. 4 was observed with mild diarrhea at 1 and 2 hours, followed by normal clinical signs till day 14. Animal no. 2 was observed with normal clinical signs at 30 minutes, mild lethargy at 1, 2, 3 and 4 hours and mild diarrhoea at 1 and 2 hours, followed by normal clinical signs till day 14. Animal no. 3 was observed with mild lethargy at 30 minutes, 1, 2, 3, 4 hours, day 1, 2 and 3 and mild diarrhoea at 2 hours, followed by normal clinical signs till day 14. Animal no. 5 was observed with mild lethargy at 30 minutes, 1, 2, 3 and 4 hours and mild diarrhoea at 1, 2, 3 and 4 hours, followed by normal clinical signs till day 14. Animal no. 6 was observed normal clinical signs at 30 minutes, 1, 2, 3 and 4 hours and mild lethargy on day 1 and 2, followed by normal clinical signs till day 14
Body weight:
Mean body weight of animals treated with 2000 mg/kg body weight was observed with gain on day 7 and 14, as compared to day 0
Gross pathology:
No external and internal gross pathological examination were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice

Any other information on results incl. tables

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day

0-7

Day

0-14

1

G1/ 2000

135

160

177

18.52

31.11

2

146

180

199

23.29

36.30

3

137

143

172

4.38

25.55

4

147

166

182

12.93

23.81

5

160

190

206

18.75

28.75

6

131

161

183

22.90

39.69

Key:- = Not Applicable

 


Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)

Sex:Female

Group/ Dose (mg/kg)

Rats Body Weight (g)

Body Weight Changes (%)

Day 0

Day 7

Day 14

0-7

0-14

 

G1/ 2000

Mean

142.67

166.67

186.50

16.79

30.87

SD

10.56

16.49

13.19

7.15

6.17

n

6

6

6

6

6

Keys:SD = Standard Deviation, n = Number of Animals

Keys:- = Not applicable, 1 = Normal, 2 = Found dead, 4 = Abdominal breathing, 145 = Salivation,      155 = Sternal recumbency, 166 = Tremors, + = Mild, ++ = Moderate



Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral LD50 (cut-off value) of Solvent blue 4 (CAS No. – 6786-83-0) was greater than 2000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that Solvent blue 4 (CAS No.- 6786-83-0) does not exhibits acute oral toxicity in as per CLP criteria of classification
Executive summary:

Acute Oral Toxicity Study ofSolvent blue 4 (CAS No. – 6786-83-0)in Rats, This study was performed as per OECD No. 423.

Six female Wistar rats were selected for acute oral toxicity study. The animals were fasted for minimum 16-18 hours prior to dosing and for 4 hours post dosing, with food withheld but drinking water providedad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs. Three rats of group G1 were dosed with starting dose of 2000 mg/kg body weight and the animals showed no mortality post dosing, so another three rats of the same group were dosed with 2000 mg/kg weight and no mortality was observed. Hence, further dosing was stopped. Body weights were re­corded on day 0 (prior to dosing) 7 and 14. Mean body weight of all six animals was observed with gain on day 7 and 14, as compared to day 0. At 2000mg/kg, animal no. 1 and 4 were observed with mild lethargy at 30 minutes, 1, 2, 3, 4 hours and on day 1 post dosing while additionally, animal no. 1 was observed with mild lethargy on day 2 and animal no. 4 was observed with mild diarrhea at 1 and 2 hours, followed by normal clinical signs till day 14. Animal no. 2 was observed with normal clinical signs at 30 minutes, mild lethargy at 1, 2, 3 and 4 hours and mild diarrhoea at 1 and 2 hours, followed by normal clinical signs till day 14. Animal no. 3 was observed with mild lethargy at 30 minutes, 1, 2, 3, 4 hours, day 1, 2 and 3 and mild diarrhoea at 2 hours, followed by normal clinical signs till day 14. Animal no. 5 was observed with mild lethargy at 30 minutes, 1, 2, 3 and 4 hours and mild diarrhoea at 1, 2, 3 and 4 hours, followed by normal clinical signs till day 14. Animal no. 6 was observed normal clinical signs at 30 minutes, 1, 2, 3 and 4 hours and mild lethargy on day 1 and 2, followed by normal clinical signs till day 14

No external and internal gross pathological examination were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifce

Under the conditions of this; acute oral toxicity study of Solvent blue 4 (CAS No. – 6786-83-0) in female rats is as given below:The acute oralLD50(Cut-off value) of Solvent blue 4 (CAS No. – 6786-83-0)was greater than 2000 mg/kgbody weight

Solvent blue 4 (CAS No. – 6786-83-0) isbeing not classified as per CLP regulation.