Trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate

Endpoint summary

Description of key information

Acute Oral:

The acute oral LD50 in male DDY mice was determined to be greater than 2000 mg/kg after dosing with Amaranth dye.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: data from peer- reviewed journals

Qualifier:

equivalent or similar to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To determine the acute oral toxicity of the Amaranth

GLP compliance:

not specified

Test type:

other: comet assay

Limit test:

yes

Species:

mouse

Strain:

other: DDY

Sex:

male

Details on test animals or test system and environmental conditions:

Details on test animal
TEST ANIMALS
Source: Japan SLC Co., Shizuoka, Japan
Age at study initiation: 7 weeks
Weight at study initiation: no data
Fasting period before study: no data
Housing: no data
Diet (e.g. ad libitum): commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan), ad libitum
Water (e.g. ad libitum): Tap Water, ad libitum
Acclimation period: 1 week of acclimatization

ENVIRONMENTAL CONDITIONS
Temperature (°C): The animal room was at 20–24°C
Humidity (%): No data
Air changes (per hr): No data
Photoperiod (hrs dark / hrs light): 12 h
Light – dark cycle.

Route of administration:

oral: unspecified

Vehicle:

physiological saline

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED:
0.5 × LD50 or the limit dose of 2000 mg/kg.

Doses:

0.5 × LD50 or the limit dose of 2000 mg/kg.

No. of animals per sex per dose:

4 - 5 male mice

Control animals:

not specified

Details on study design:

no data

Statistics:

no data

Preliminary study:

no data

Sex:

male

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

no mortality observed

Clinical signs:

other: no data

Gross pathology:

no data

Other findings:

no data

Table 1: Acute Oral Toxicity study results

 

Test chemical	Vehicle	Source*	LD50** mg/kg
Amaranth	Saline	T	>2000

 

*T = Tokyo Kasei Kogyo Industry Ltd., Tokyo, Japan;

** In order to set appropriate doses for the assay, we determined approximate LD50 by simple acute toxicity experiments on four–five animals. When no death was observed at 2000 mg/kg, the LD50 was defined as >2000 mg/kg.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

A simple acute oral toxicity experiment was conducted prior to Comet assay in male DDY mice.

When no death was observed at 2000 mg/kg, the LD50 was defined as >2000 mg/kg.
The acute oral LD50 in male DDY mice was determined to be greater than 2000 mg/kg after dosing with Amaranth

Executive summary:

A simple acute oral toxicity experiment was conducted prior to Comet assay in male DDY mice.

Male ddY mice were obtained from Japan SLC Co., Shizuoka, Japan, at 7 weeks of age and used after 1 week of acclimatization. They were fed commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan) and tap water ad libitum throughout the acclimatization period and the experiment. The animal room was at 20–24◦C with a 12 h light – dark cycle.

 

In order to set appropriate doses for the assay, we determined approximate LD50 by simple acute toxicity experiments on four–five animals. When no death was observed at 2000 mg/kg, the LD50 was defined as >2000 mg/kg.

 

The acute oral LD50 in male DDY mice was determined to be greater than 2000 mg/kg after dosing with Amaranth dye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Simple acute oral toxicity experiment was conducted prior to Comet assay in male DDY mice. When no death was observed at 2000 mg/kg, the LD50 was defined as >2000 mg/kg. The acute oral LD50 in male DDY mice was determined to be greater than 2000 mg/kg after dosing with Amaranth

Additional information

Acute oral toxicity:

Peer reviewed articles were viewed to determine the acute oral toxic nature of the test compound trisodium (4E)-3-oxo-4-[(4- sulfonato-1- naphthyl)hydrazono]naphthalene- 2,7-disulfonate (CAS no 915-67-3). The studies are summarized as below:

 

A simple acute oral toxicity (The comet assay with 8 mouse organs: results with 39 currently used food additives, 2002) experiment was conducted prior to Comet assay in male DDY mice.

Male ddY mice were obtained from Japan SLC Co., Shizuoka, Japan, at 7 weeks of age and used after 1 week of acclimatization. They were fed commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan) and tap water ad libitum throughout the acclimatization period and the experiment. The animal room was at 20–24°C with a 12 h light – dark cycle. In order to set appropriate doses for the assay, we determined approximate LD50 by simple acute toxicity experiments on four–five animals. When no death was observed at 2000 mg/kg, the LD50 was defined as >2000 mg/kg.

The acute oral LD50 in male DDY mice was determined to be greater than 2000 mg/kg after dosing with Amaranth dye.

 

A simple acute oral toxicity experiment was conducted prior to DNA damage in pregnant female CD1 mice. Samples of amaranth (Food Red No. 2, CAS (915–67–3), was from certified lots approved for food use in Japan and was purchased from Tokyo Kasei Organic Chemicals (Tokyo).

CD-1 (ICR) mice were obtained from Charles River Japan, Inc. (Yokohama) at 7 weeks of age and used for the experiments after 1 week of acclimatization. They were fed with commercial pellets MF (Oriental Yeast Industries, Tokyo) and tap waterad libitumthroughout the acclimatization period and the experiment. The animal room was kept at 22 ± 2°C with a 12-h light-dark cycle; the humidity was 30–50%. For the pregnant mouse study, female mice were mated for a period of one to two days. The morning on which copulation plugs were observed was designated day 0 of gestation. On the morning of day 11, 4 mice were randomly assigned to each treatment group. Chemicals were dissolved in distilled water and administered by gavage at the limit dose of 2000 mg/kg (10 ml/kg). The same volume of distilled water was administered to the control mice at the same time. The limit dose, at which no deaths, morbidity, or distinctive clinical signs were observed, was determined by preliminary acute toxicity tests. The Limit dose at which no mortality, morbidity or toxic signs were observed in mice dosed with Amaranth was determined to be greater than 2000 mg/kg

 

From Scientific Opinion on the re-evaluation of Amaranth (E 123) as a food additive (2010), acute Oral toxicity studies were carried out in mice. Amaranth was orally dosed in mice and toxic effects were observed for 14 days. The Acute Oral LD50 for Amaranth Dye was reported to be greater than 10000mg/kgb.w in mice.

 

From the same study, Acute Oral toxicity studies were carried out in rats to determine the toxicity of Amaranth. Amaranth was dosed orally in rats and toxic signs were observed for 14 days.  The Acute Oral LD50 for Amaranth Dye was reported to be 6000mg/kgb.w in rats.

 

From the SSS qsar prediction, The acute median lethal dose (LD50) value of2,7-Naphthalenedisulfonic acid, 3-hydroxy-4-[(4-sulfo-1-naphthalenyl), sodium salt (CAS No 915-67-3)in rat is estimated to be 6901.0312 mg/kg bw. This value indicates that the substance is not expected to exhibit acute toxicity by oral route as per the CLP criteria.

 

From Differential colon DNA damage induced by azo food additives between rats and mice (2010) study, a simple acute oral toxicity experiment was conducted prior to Comet assay in male ICR - CD1 mice.

Samples of Amaranth were from certified lots for use in foods and were purchased from Tokyo Chemical Industry Limited Co., Japan. Male ICR –CD1 mice were obtained from Charles River Japan, Inc, Japan, at 7 weeks of age and used after 1 week of acclimatization. They were fed commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan) and tap water ad libitum throughout the acclimatization period and the experiment. The animal room was at 22–24◦C, and 50 -70% humidity with a 12 h light – dark cycle. Amaranth dye was dissolved in distilled water and administered by gavage in a volume of 10ml/kg. Each animal was administered the test solution at 10 to 11 a.m and was fed with commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan) and tap water ad libitum throughout the study duration. Four mice were assigned to each study group (1 and 10 mg/kg). 6 mice dosed with distilled water only acted as controls. Shortly after dosing and before sacrifice the test animals were carefully observed for pharmacotoxic signs. Mice were sacrificed by cervical dislocation 3 hours after treatment with the test chemical. No death, distinctive clinical signs, mortality or morbidity were observed after treatment. Necropsy revealed no toxic effects on all organs. The acute oral LD50 in male ICR –CD1 mice was determined to be greater than 10 mg/kg after dosing with Amaranth.

 

From the same study, A simple acute oral toxicity experiment was conducted prior to Comet assay in male Fisher 344 rats. Samples of Amaranth were from certified lots for use in foods and were purchased from Tokyo Chemical Industry Limited Co., Japan. Male Fischer 344 rats were obtained from Charles River Japan, Inc, Japan, at 7 weeks of age and used after 1 week of acclimatization. They were fed commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan) and tap water ad libitum throughout the acclimatization period and the experiment. The animal room was at 22–24◦C, and 50 -70% humidity with a 12 h light – dark cycle. Amaranth dye was dissolved in distilled water and administered by gavage in a volume of 10ml/kg. Each animal was administered the test solution at 10 to 11 a.m and was fed with commercial pellets MF (Oriental Yeast Industries Co., Tokyo, Japan) and tap water ad libitum throughout the study duration. Four – five rats were assigned to each study group (10, 100, 1000 mg/kg). 5 rats dosed with distilled water only acted as controls. Shortly after dosing and before sacrifice the test animals were carefully observed for pharmacotoxic signs. Rats were sacrificed by anesthesia with ether 3 hours after treatment of amaranth (10 mg/kg) and 24 hours after treatment of amaranth (100, 1000 mg/kg) and necropsied. No death, distinctive clinical signs, mortality or morbidity were observed after treatment. Necropsy revealed no toxic effects on all organs. The acute oral LD50 in male Fischer 344 rats was determined to be greater than 1000 mg/kg after dosing with Amaranth.

 

Based on the majority of studies reviewed and as per the CLP classification, the test material Amaranth (915-67-3) does not classify as an acute oral toxicant.

Justification for selection of acute toxicity – oral endpoint
Data from peer- reviewed journals

Justification for classification or non-classification

Acute oral toxicity:

Based on the studies reviewed and as per the CLP classification, the test material Amaranth (915-67-3) does not classify as an acute oral toxicant.