[No public or meaningful name is available]

Endpoint summary

Description of key information

LD₅₀ (oral, rat, female) > 2000 mg/kg bw

LD₅₀ (dermal, rat, 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 July 21 to August 11, 2005

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: 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 III suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: certified rat and mouse diet (Code 5LF2) supplied by BCM IPS Limited, ad libitum.
- Water: drinking water, ad libitum.
- Acclimation period: at least five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 19 to 25 °C
- Relative humidity: 30 to 70 %
- Air changes: 15 changes per hour.
- Photoperiod: 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:

For the purpose of the study the test material was freshly prepared, as required, as a solution at the appropriate concentration in distilled water.

Sufficient time was allowed between each group to confirm the survival of the previously dosed animals.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 rats × group

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: the animals were observed for deaths or overt signs of toxicity 30 min, 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: yes; 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. Black faeces were noted in three animals one and two days after dosing.

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 (females) > 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 guideline 423 and EU Method B1 tris Acute Toxicity (Oral) of Commission Directive 2004173/EC Method. 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 and there were no signs of systemic toxicity. All animals showed expected gains in bodyweight over the study period.

No abnormalities were noted at necropsy.

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.

Conclusion

LD50 (females) > 2000 mg/kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

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 October 18 to November 01, 1990

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:

Details for read across approach are included into 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: BRL Ltd., Basel, Switzerland.
- Age at study initiation: approx. 11 weeks.
- Weight at study initiation: males 309 to 322 g and females 202 to 240 g
- Housing: individually housed in polycarbonate cages containing purified sawdust as bedding material (Woody SPF, supplied by Broekman Institute, Someren, The Netherlands).
- Diet: free access to standard pelleted laboratory animal diet (Kliba 343 from Klingentalmdhle AG, Kaiseraugst, Switzerland).
- Water: free access to tap-water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 °C
- Humidity: 55 %:
- Air changes: 15 air changes per hour.
- Photoperiod: lighting was 12 hours artificial fluorescent light and 12 hours dark per day.

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: one day before exposure an area of approximately 5×7 cm on the back of the animal was clipped. The formulation was applied to an area of approximately 25 cm2 (5×5 cm) for males and 18 cm2 (3.5×5 cm) for females by application on a gauze patch.
- Type of wrap if used: gauze patch was fixed to aluminium foil end flexible bandage, with drops of petrolatum.

REMOVAL OF TEST SUBSTANCE
- Washing: whereafter residual test substance was removed with tissue moistened with tap-water.
- Time after start of exposure: 24 hours.

TEST SUBSTANCE PREPARATION
The formulations were prepared immediately prior to dosing. The test substance was weighed into a glass flask on an analytical balance and the vehicle (wlw) was added. Following stirring the test substance preparation formed a solution.

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
- Mortality / Viability: at periodic intervals on the day of dosing (day 1) and twice daily thereafter for 14 days.
- Body Weights: days 1 (pre-administration), 8 and 15.
- Symptoms: at periodic intervals on the day of dosing (day 1) and once daily thereafter For 14 days. All signs of reaction to treatment were recorded with particular attention paid to changes in the skin (treated skin), fur, eyes and mucous membranes, as well as to behaviour pattern, tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Necropsy of survivors performed: all animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide asphyxiation and subjected to necropsy. All animals assigned to the study were subjected to necropsy. The necropsies were performed by experienced prosectors and descriptions of all macroscopic abnormalities recorded.

Sex:

male/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:

No mortality occurred during the study period.

Clinical signs:

No clinical signs of ill health or behavioural changes were seen during the study.
No skin irritation was observed on the treated area during the study period. All animals showed blue staining of the treated skin area by the test substance during the study period.

Body weight:

Slight body weight gain was noted among all animals over the first week of observation. All animals showed improved body weight gain over the second week of the study.

Gross pathology:

Macroscopic post mortem examination of the surviving animals at termination did not reveal any abnormalities.

Interpretation of results:

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

Conclusions:

LD50 (males and females) > 2000 mg/kg bw

Executive summary:

The purpose of the study was to assess the toxicity of the test article when administered to rats as a single dermal dose. The study was carried out in accordance with OECD guideline 402 and EEC Directive 84/449/EEC, Part B.3, "Acute Toxicity ~ Dermal". Test item was administered by dermal application, to five rats of each sex, at 2000 mg/kg body weight. Macroscopic examination was performed at the end of the experimental period.

No animals died during the study. No clinical signs of ill health or behavioural changes were seen during the observation period. Slight body weight gain was noted over the first week with recovery of body weight gain over the second week of observation.

No signs of skin irritation were observed on the treated area during the study period. All animals showed blue staining of the treated skin by the test substance.

Macroscopic post mortem examination of the surviving animals at termination did not reveal any abnormalities.

Conclusion

LD50 (males and females) > 2000 mg/kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

ORAL ACUTE TOXICITY

The acute oral toxicity potential of Direct Blue 267 was investigated in rats. The method was designed to meet the requirements of the OECD Guidelines 423 and EU Method B1 tris. Animals were treated with the test material at a dose level of 2000 mg/kg bodyweight. No deaths and no signs of systemic toxicity were recorded. All animals showed expected gains in bodyweight over the study period. No abnormalities were noted at necropsy. Therefore, 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 (Mullaney, 2005).

INHALATION ACUTE TOXICITY

No acute toxicity studies by inhalation route are available on Direct Blue 267. However, due to the physical state and the trade forms of the substance, inhalation is not an appropriate route of exposure. In addition, the substance is manufactured and handled with suitable risk management measures and with the suitable personal protective equipments; the use consitions lets to consider the possible absorption of the substance by inhalation route as negligible.

DERMAL ACUTE TOXICITY

According to the Commission Regulation (EU) 2016/863, testing by dermal route does not need to be conducted if no systemic effects have been observed in in vivo studies with dermal exposure. Furthermore, recent scientific analysis of available data from in vivo acute toxicity studies have shown that substances that are not toxic via oral route may be expected with high certainty to be also non-toxic via dermal route. Therefore, testing those substances via dermal route does not provide essential information for their safety assessment.

The amendment is a 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 was 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 up to 2000 mg/kg bw.

The expectation is confirmed by available data Similar Substance 01; the read across approach can be considered reliable and appropriate to investigate the property (details for the approach are included into the IUCLID section 13).

The purpose of the study was to assess the toxicity of the Similar Substance 01 when administered to rats as a single dermal dose. The study was carried out in accordance with OECD guideline 402. Test item was administered by dermal application to five rats of each sex, at 2000 mg/kg body weight. No animals died during the study. No clinical signs of ill health or behavioural changes were seen during the observation period. Slight body weight gain was noted over the first week with recovery of body weight gain over the second week of observation. No signs of skin irritation were observed on the treated area during the study period. All animals showed blue staining of the treated skin by the test substance. Macroscopic post mortem examination of the surviving animals at termination did not reveal any abnormalities (Reijnders, 1991).

REFERENCE

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 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by 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 test substance is out of the 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 the 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, test substance does non meet the criteria to be classified for either oral, neither dermal acute toxicity, according to the CLP Regulation.