Sodium 1-amino-4-[[3,5-bis[[(chloroacetyl)amino]methyl]-2,4,6-trimethylphenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Endpoint summary

Description of key information

Acid Blue 225 is found to have low acute toxicity with oral LD50 >6000 mg/kg bw.

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:

14 March 1974

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

Name: 21036 / A
Purity: 79.9 %

Species:

rat

Strain:

other: Tif. RAI

Sex:

male/female

Details on test animals or test system and environmental conditions:

The compound was tested on 40 Tif. .RAI rats (20 males/20 females), bred under SPF conditions in our own breeding unit. They were 6 to 7 weeks old and weighed 160 to 180 g. The males and females were segregated and housed in Macrolon cages (Type 3) in groups of 5 in a room kept at a constant temperature of 22 ± 1 °C and a relative humidity of approximately 50 %. They received water and food (NAFAG, Gossau SG, rat food) ad libitum. The rats were starved during one night before starting the treatment.

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Doses:

1000, 3170, 4640 and 6000 mg/kg

No. of animals per sex per dose:

5 male and 5 females

Control animals:

no

Details on study design:

FAT 21036/A was weighed into an Erlenmeyer flask on a Mettler balance. It was suspended at 10 and 30 % with carboxymethylcellulose 2 % and administered by oral intubation. Before treatment the suspension was homogeneously dispersed with an Ultra-Turrax and during treatment it was kept stable with a magnetic stirrer.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 6 000 mg/kg bw

Based on:

test mat.

Mortality:

One animal was found dead in 6000 mg/kg group.

Clinical signs:

other: Within 2 hours after treatment the rats in all dosage groups showed sedation, dyspnoea, exophthalmos, curved position and ruffled fur. These symptoms became more accentuated as the dose was increased. The surviving animals had recovered within 4 to 6 days

Gross pathology:

No substance related gross organ changes were seen.

Other findings:

No further abnormalities reported.

Mortality:

Dose mg/kg	Concentration % of formulation	No of Animals		Died within
		m	f	m 2hr	f 1hr	m 24 hrs	f24 hrs	m 48 hrs	f 48 hrs	m 7 d	f  7 d	m 14 d	f 14 d
1000	10	5	5	0	0	0	0	0	0	0	0	0	0
3170	30	5	5	0	0	0	0	0	0	0	0	0	0
4640	30	5	5	0	0	0	0	0	0	0	0	0	0
6000	30	5	5	0	1	0	1	1	1	1	1	1	1
No higher dosed were possible

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral LD50 of FAT 21036/A in rats of both sexes observed over a period of 7 days is greater than 6000 mg/kg.

Executive summary:

The acute oral toxicity of FAT 21036/A was assessed using Tif. RAI rats in a study conducted using methodology similar to OECD Guideline 401. FAT 21036/A was suspended at 30 % with polyethylene glycol (PEG 400). The test material was administered at different doses (5 males and 5 females rats per dose): 1000, 3170, 4640 and 6000 mg/kg. Within 2 hours after treatment the rats in all dosage groups showed sedation, dyspnoea, exophthalmos, curved position and ruffled fur. These symptoms became more accentuated as the dose was increased. The surviving animals had recovered within 4 to 6 days. No substance related gross organ changes were seen at the post mortem examination. In conclusion, the acute oral LD50 of FAT 21036/A in rats of both sexes observed over a period of 7 days is greater than 6000 mg/kg. The compound therefore is non-toxic to the rat by this route of administration.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

6 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute toxicity: oral

The acute oral toxicity of FAT 21036/A was assessed using Tif. RAI rats in a study conducted using methodology similar to OECD Guideline 401. FAT 21036/A was suspended at 30 % with polyethylene glycol (PEG 400). The test material was administered at different doses (5 males and 5 females rats per dose): 1000, 3170, 4640 and 6000 mg/kg. Within 2 hours after treatment the rats in all dosage groups showed sedation, dyspnoea, exophthalmos, curved position and ruffled fur. These symptoms became more accentuated as the dose was increased. The surviving animals had recovered within 4 to 6 days. No substance related gross organ changes were seen at the post mortem examination. In conclusion, the acute oral LD50 of FAT 21036/A in rats of both sexes observed over a period of 7 days is greater than 6000 mg/kg. The compound therefore is non-toxic to the rat by this route of administration.

 

Acute toxicity: inhalation

Currently no study to assess the acute inhalation toxicity potential of Acid Blue 225 is available. However, the vapour pressure for the substance can be considered low owing to the high melting point (>300 °C). Hence, the substance is considered to have low volatility. Synthesis and formulation of this chemical is performed in a closed process; the final product consists of liquid formulations only. Hence, the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Based on column 2, ‘Specific rules for adaptation from column 1’ of the table given in REACH Annex VII, the study on acute inhalation toxicity only needs to be conducted if an exposure via inhalation is to be expected, based on vapour pressure and/or the likelihood of an exposure to aerosols, particles or droplets. Referring to the expected low volatility of the substance, the fact that the substance is imported into the EU in a formulated form as liquid formulation, the exposure via inhalation is considered to be unlikely. Further, in case the substance enters the respiratory tract, it will be trapped in the mucus and cleared owing to its high water solubility of 64.3 g/L, thereby further limiting the absorption. The chemical showed low toxicity potential in the available acute oral toxicity study (LD50 >6000 mg/kg bw) with no mortality or systemic toxicity up to 2000 mg/kg bw, hence it does not need to be classified STOT SE. Taking the above arguments into account, low toxicity potential is expected on acute exposure of Acid Blue 225 via inhalation route and hence testing by the inhalation route was considered scientifically not necessary.

 

Acute toxicity: dermal

Currently no study to assess the acute dermal toxicity potential of Acid Blue 225 is available. However, the molecular weight of Acid Blue 225 is 669.5 g/mol, indicating it being too large for dermal absorption.It has water solubility of 64.3 g/L and n-octanol/water partition coefficient (log P) of 0.58, indicating it being too hydrophilic to cross the lipid rich environment of the stratum corneum. Hence, the dermal uptake for the substance will be low.The chemical showed low toxicity potential in the available acute oral toxicity study (LD₅₀>6000 mg/kg bw), with no mortality or systemic toxicity being seen up to 2000 mg/kg bw, hence it does not need to be classified as STOT SE. Similarly, absence of systemic toxicity in skin irritation and sensitisation studies further supports the conclusion that no adverse effects are expected for the chemical via the dermal route. Further, experience with similar chemical substances has demonstrated that it is very unlikely that toxicity related to the intrinsic properties of the chemical only show up upon dermal exposure and not after systemic application. Taking above arguments into account, low toxicity is expected on acute dermal exposure of Acid Blue 225 and testing by the dermal route was considered scientifically not necessary.

Justification for classification or non-classification

Acid Blue 225 is found to have low acute toxicity with oral LD50 >6000 mg/kg bw, hence it does not meet the criteria of classification for acute toxicty and/or specific target organ toxicity on single exposure according to the Regulation (EC) No. 1272/2008.