Disodium 4-amino-3-[[4-[(2,4-diaminophenyl)azo]phenyl]azo]-5-hydroxy-6-(phenylazo)naphthalene-2,7-disulphonate

Endpoint summary

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

2001

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 12 weeks of age
- Weight at study initiation:
- Fasting period before study: overnight
- Housing: In groups of up to four in suspended solid-floor polypropylene cages furnished with softwood flake bedding.
- Diet: free access to 2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK
- Water: free access
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL and 200 mg/mL
- Amount of vehicle (if gavage): not specified
- Justification for choice of vehicle: solubility of the test item

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.

Doses:

In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose and a single animal was treated.
In the absence of toxicity at a dose level of 300 mg/kg, an additional animal was treated at a dose level of 2000 mg/kg.
In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of 4 animals was treated at a dose level of 2000 mg/kg.

No. of animals per sex per dose:

1 female, 300 mg/kg
5 female, 2000 mg/kg

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays. Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

Dose Level - 300 mg/kg: there were no deaths
Dose Level - 2000 mg/kg : There were no deaths.

Clinical signs:

other: Dose Level - 300 mg/kg: No signs of systemic toxicity were noted during the observation period Dose Level - 2000 mg/kg : No signs of systemic toxicity were noted during the observation period. Black stained feces were noted in one treated animal one to fo

Gross pathology:

Dose Level - 300 mg/kg: No abnormalities were noted at necropsy.
Dose Level - 2000 mg/kg : Patchy pallor of the liver was noted in one treated animal. No abnormalities were noted at the necropsy of the four other animals.

Appendix 1 Individual Clinical Observations and Mortality Data - 300 mg/kg

Dose level (mg/kg)	Animal number and sex	Effects noted after dosing (hours)				Effects noted during period after dosing (days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
300	1-0 female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Appendix 2 Individual Body Weights and Body Weight Changes - 300 mg/kg

Dose level (mg/kg)	Animal number and sex	Body Weight (g) at Day			Body Weight Gain (g) During Week
		0	7	14	1	2
300	1-0 female	208	212	221	4	9

Appendix 3 Individual Necropsy Findings - 300 mg/kg

Dose Level (mg/kg)	Animal Number and Sex	Time of Death	Macroscopic Observations
300	1-0 female	Killed Day 14	No abnormalities detected

Appendix 4 Individual Clinical Observations and Mortality Data - 2000 mg/kg

Dose level (mg/kg)	Animal number and sex	Effects noted after dosing (hours)				Effects noted during period after dosing (days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	2-0 female	0	0	0	0	F0	F0	F0	F0	0	0	0	0	0	0	0	0	0	0
	3-0 female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-1 female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-2 female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	3-3 female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

                                    

0 = No signs of systemic toxicity ; F = Feces stained black

Appendix 5 Individual Body Weights and Body Weight Changes - 2000 mg/kg

Dose level (mg/kg)	Animal number and sex	Body Weight (g) at Day			Body Weight Gain (g) During Week
		0	7	14	1	2
2000	2-0 female	221	221	234	0	13
	3-0 female	209	220	233	11	13
	3-1 female	203	219	229	16	10
	3-2 female	202	228	238	26	10
	3-3 female	208	219	227	11	8

 

Appendix 6 Individual Necropsy Findings - 2000 mg/kg

Dose Level (mg/kg)	Animal Number and Sex	Time of Death	Macroscopic Observations
2000	2-0 female	Killed Day 14	Liver: Patchy pallor
	3-0 female	Killed Day 14	No abnormalities detected
	3-1 female	Killed Day 14	No abnormalities detected
	3-2 female	Killed Day 14	No abnormalities detected
	3-3 female	Killed Day 14	No abnormalities detected

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight.

Executive summary:

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat. Following a sighting test at dose levels of 300 mg/kg and 2000 mg/kg, a further group of four fasted females was given a single oral dose of test item, as a solution in distilled water, at a dose level of 2000 mg/kg body weight.  Clinical signs and body weight development were monitored during the study.  All animals were subjected to gross necropsy.

There were no deaths. There were no signs of systemic toxicity. All animals showed expected gains in body weight. Patchy pallor of the liver was noted at necropsy of one animal treated at a dose level of 2000 mg/kg.  No other abnormalities were noted.

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System − Unclassified).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Quality of whole database:

Results of a study following OECD Guideline 420, GLP compliant.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The result of one Acute Oral Toxicity study, conducted according to the OECD 420 guideline, gave as a result that the acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be > 2000 mg/kg body weight. According to the criteria described in the Annex I of the EC Regulation 1272/2008, the substance is not classified as Acute Tox Oral.