Disodium 3,3'-[[6-[bis(2-hydroxyethyl)amino]-1,3,5-triazine-2,4-diyl]bis[imino(3-methoxy-4,1-phenylene)azo]]bis[benzenesulphonate]

Endpoint summary

Description of key information

Not harmful/toxic if swallowed (oral LD50 (female) > 2000 mg/kg bw)

Not harmful/toxic in contact with skin (dermal LD50 (male and female) > 2000 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:

From May 18 to June 08, 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 17 December 2001

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK.
- Females: animals were nulliparous and non-pregnant.
- Age at study initiation: eight to twelve weeks of age.
- Weight at study initiation: bodyweights fell within an interval of ± 20 % of the mean initial bodyweight of the first treated group.
- Fasting period before study: overnight fast immediately before dosing and for approximately three to four hours after dosing.
- Housing: animals were housed in groups of three In suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: Certified Rat and Mouse Diet (Code 5LF2) supplied by BCM IPS Limited, ad libitum.
- Wate: ad libitum.
- Acclimation period: at least five days.

FOOD AND WATER QUALITY
The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature: 19 - 25 °C
- Humidity: 30 - 70 %
- Air changes: at least fifteen changes per hour.
- Photoperiod: the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

- Concentration: 200 mg/ml
- Volume: 10 mg/kg

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Three rats

Details on study design:

- Procedure: treatment of animals was sequential. Sufficient time was allowed between each group to confirm the survival of the previously dosed animals.
- Duration of observation period following administration: 14 days
- Frequency of observations: the animals were observed for deaths or overt signs of toxicity 30 minutes, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.
- Frequency of weighing: individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment.
- Necropsy of survivors performed: at the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

There were no deaths.

Clinical signs:

There were no signs of systemic toxicity.

Body weight:

All animals showed expected gains in bodyweight over the study period.

Gross pathology:

No abnormalities were noted at necropsy.

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

LD50 (female) > 2000 mg/kg bw

Executive summary:

The study was performed to assess the acute oral toxicity of the test material following a single oral administration in the Sprague-Dawley CD strain rat. The method was designed to meet the requirements of the OECD Guidelines 423 and the EU Method B1 tris A group of three fasted females was treated with the test material at a dose level of 2000 mg/kg bodyweight. This was followed by a further group of three fasted females at the same dose level.

The test material was administered orally as a solution in distilled water. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

There were no deaths during the experiment; no signs of systemic toxicity was recorded. All animals showed expected gains in bodyweight over the study period.

No abnormalities were noted at necropsy.

Conclusion

The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From November 18 to December 07, 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

The read across approach has been detailed in the report attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted February 24, 1987

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, West-Germany.
- Age at study initiation: 9 weeks.
- Weight at study initiation: 308 - 377 g males; 229 - 234 g females.
- Housing: individually housed in polycarbonate cages containing purified sawdust as bedding material (Woody Clean supplied by Broekman Institute, Someren, The Netherlands).
- Diet: standard pelleted laboratory animal diet (RMH-B from Hope Farms, Woerden, The Netherlands), ad libitum.
- Water: tap-water, diluted with decalcified water, ad libitum.
- Acclimation period: 5 days under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 3 °C
- Relative dumidity: 30 - 70 %
- Air changes: 7.5-15 air changes per hour.
- Photoperiod: 12 hours artificial fluorescent light/12 hours dark per day.

Vehicle:

water

Remarks:

prepared by reverse osmosis

Details on dermal exposure:

FORMULATION
The formulations were prepared immediately prior to dosing. The test article was weighed into a glass flask on an analytical balance and the vehicle (w/w) was added.
Homogeneity of the test article in vehicle was obtained by a homogeniser.

TREATMENT
- Shaving: one day before treatment (day -1) an area of approximately 5 x 7 cm on the back of the animal was clipped.
- Application: approximately 25 cm^2 (5 x 5 cm) for males and 18 cm2 (3.5 x 5 cm) for females was applied using a gauze patch fixed, successively, to aluminium foil and flexible bandage (Cohan, 3M, St. Paul, U.S.A.), with drops of vaseline.
- Dose volume: 10 ml/kg bw

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations: at periodic intervals on the day dosing (day 1) and twice daily thereafter for at least 14 days. All signs of reaction to treatment were recorded with particular attention paid to changes in the skin, fur, eyes and mucous membranes, as well as to behaviour pattern, tremors, convuision, salivations, diarrhoea, lethargy, sleep and coma. Changes of the treated skin were described immediately after bandage removal (day 2), on day 5, 8 and 15.
- Frequency of weighing: test days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: all animals were necropsied and descriptions of all macroscopic abnormalities were recorded. All animals surviving to the end of the observation period were sacrified by carbon dioxide asphyxiation and subjected to necropsy.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality occurred during the study period.

Clinical signs:

There were no treatment related signs of toxicity over the 15 day observation period.
The treated skin surface of animals showed no irritation. During the study period all animals showed orange discolouration of the treated skin, due to staining by the test substance.

Body weight:

With the exception of animal 9, all animals showed body weight gain during the study period.

Gross pathology:

Macroscopic examination of all animals at the end of the study showed in one female red discolouration and thickened wall of the forestomach accompanied by blister-like areas.

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

LD50 (male and female) > 2000 mg/kg bw

Executive summary:

The substance was administered to rats of both sexes by dermal application at 2000 mg/kg body weight for 24 hours. No mortality occurred during the study period.

There were no treatment related signs of toxicity over the 15 day observation period. With the exception of animal 9, all animals showed body weight gain during the study period. The treated skin surface of animals showed no irritation. During the study period all animals showed orange discolouration of the treated skin, due to staining by the test substance. Macroscopic examination of all animals at the end of the study showed in one female red discolouration and thickened wall of the forestomach accompanied by blister-like areas.

The dermal LD50 value in rats of both sexes was estimated to exceed 2000 mg/kg body weight.

Conclusion

LD50 (male and female) > 2000 mg/kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

ORAL ACUTE TOXICITY

The acute oral toxicity of the test material was tested following a single oral administration in the Sprague-Dawley CD strain rat. The method was designed to meet the requirements of the OECD Guidelines 423 and the EU Method B1 tris. A group of three fasted females was treated with the test material at a dose level of 2000 mg/kg bodyweight. This was followed by a further group of three fasted females at the same dose level. There were no deaths during the experiment; no signs of systemic toxicity was recorded. All animals showed expected gains in bodyweight over the study period. No abnormalities were noted at necropsy.

INHALATION ACUTE TOXICITY

No acute toxicity studies by inhalation route are available on Direct Yellow 142.

Nevertheless, because of the physical state of the substance inhalation is not an appropriate route of exposure. Particle size distribution showed that the 90 % of particles has a diameter lower than ca 740 μm and that less than 10 % of particles have a diameter lower than 4 μm. Thus, the most of the Direct Yellow 142 particles are expected to remain in the upper respiratory tract, which is characterized by efficacious defence mechanisms able to remove them; therefore, respirable particles, able to enter the deepest part of the lung, the non-ciliated alveoli (i.e. D50 of 4 μm), can be considered a minimal portion.

In addition, it should be considered that the used powder form of Direct Yellow 142 contains small amount of oil, in order to avoid dusts formation and in order to facilitate the substance handling.

These considerations, together with the one that the substance is manufactured and handled with suitable risk management measures and with the suitable personal protective equipments, lets to consider the possible absorption of the substance by inhalation route as negligible.

DERMAL ACUTE TOXICITY

The inhalation and the skin contact of Direct Yellow 142 are unlikely. According to the Commission Regulation (EU) 2016/863, amending Annexes VII and VIII to REACH Regulation (EC) No 1907/2006 as regards skin corrosion/irritation, serious eye damage/eye irritation and acute toxicity, testing by the dermal route does not need to be conducted if no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation). Furthermore, it is explained that recent scientific analysis of available data from in vivo acute toxicity studies have shown that substances that are not toxic via the oral route may be expected with high certainty to be also non-toxic via the dermal route. Therefore, testing those substances via the dermal route does not provide essential information for their safety assessment. The amendment is the consequence of studies and scientific debates. In particular, the 15th Meeting of Competent Authorities for REACH and CLP (CARACAL, 2014) concluded that an adaptation of point 8.5.3 of Annex VIII to REACH is justified in order to not require information on acute dermal toxicity for substances that have shown no toxicity in acute oral toxicity test up to the limit dose of 2000 mg/kg bw. In the oral acute toxicity test, no signs of systemic toxicity were recorded. Furthermore, no reason of concern is raised on the basis of the skin/eye irritation investigations.

The expectation of non-concern for dermal acute toxicity is supported by the experimental results obtained in a test conducted on the structural analogous Similar Substance 01. The read across approach can be considered as reliable and adequate for the purpose; details and explanations are detailed in the report attached to the IUCLID section 13.

The Similar Substance 01 was administered to rats of both sexes by dermal application at 2000 mg/kg body weight for 24 hours. No mortality occurred during the study period. There were no treatment related signs of toxicity over the 15 day observation period. With the exception of animal 9, all animals showed body weight gain during the study period. The treated skin surface of animals showed no irritation. During the study period all animals showed orange discolouration of the treated skin, due to staining by the test substance. Macroscopic examination of all animals at the end of the study showed in one female red discolouration and thickened wall of the forestomach accompanied by blister-like areas.

REFERRENCE

CARACAL, 2014. 15th Meeting of Competent Authorities for REACH and CLP (CARACAL), 8 – 9 July 2014. Charlemagne building, Brussels, Belgium. Brussels, 26 July 2014. Doc. CA/61/2014. Stakeholder proposal to modify REACH standard information requirements for acute toxicity (REACH Annex VIII, point 8.5)

Justification for classification or non-classification

According to the CLP Regulation (EC) No 1272/2008, 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The oral LD50 value was established to be greater than 2000 mg/kg body weight, therefore the test substance is out of any classification limit for acute oral toxicity (oral acute toxicity category 4: 300 < ATE ≤ 2000 mg/kg bw).

The dermal LD50 value was established to be greater than 2000 mg/kg body weight, therefore the test substance is out of any classification limit for acute dermal toxicity (dermal acute toxicity category 4: 1000 < ATE ≤ 2000 mg/kg bw).

Inhalation exposure is unlikely, thus no acute toxicity value is available and no further investigation is required.

In conclusion, the test substance does not meet the criteria to be classified for oral/dermal acute toxicity, according to the CLP Regulation (EC) No 1272/2008.