Pentasodium 4-amino-6-[[5-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-2-sulphonatophenyl]azo]-3-[(2,5-disulphonatophenyl)azo]-5-hydroxynaphthalene-2,7-disulphonate

Endpoint summary

Description of key information

Reactive Black 039 was found to have an oral LD50 >5000 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 17 September, 1992 to 08 October, 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Principles of method if other than guideline:

Guidelines followed

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: HanIbm: WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: and BRL, Biological Research Laboratories, Ltd. Wolferstrasse 4, CH-4414 Füllinsdorf
- Age at study initiation: males: 8 weeks; females: 10 weeks
- Weight at study initiation: males: 190-205 g; females: 172-182 g
- Housing: Groups of five in Makrolon type-3 cages (size: 22 x 37.5 x 15 cm) with standard (softwood bedding ("Lignocel" , Schill AG, CH-4132 Muttenz).
- Diet: Pelleted standard Kliba 343, Batch 88/92 rat maintenance diet ("Kliba". Klingentalmuehle AG. CH-4303 Kaiseraugst) (ad libitum except for overnight fasting period)
- Water: Community tap water ad libitum
- Acclimation period: One week under laboratory conditions, after veterinary examination. Only animals without any visual signs of illness were used for the study.
- Identification: by unique cage number and corresponding color-coded spots on the tail
- Randomization: Randomly selected at time of delivery in groups of five.
- Fasting period before study: 16 to 17 h

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 40-70 %
- Air changes: 10-15 air changes/h
- Photoperiod: 12 h/12 h (music during the light period)

IN-LIFE DATES: From: September 17, 1992; To: October 8, 1992

Route of administration:

oral: gavage

Vehicle:

other: bidistilled water

Details on oral exposure:

TEST SUBSTANCE PREPARATION
The test substance was placed into a glass beaker on a tared Mettler PM 480 balance, and the vehicle (bidistilled water) was added. A w/v dilution was prepared using a homogenizer. Homogeneity of the test substance in the vehicle was maintained during treatment using a magnetic stirrer. The preparation was made immediately prior to each dosing.
Application Volume/kg bw: 10 mL at 2000 mg/kg bw

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5/sex/dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 15 d
- Necropsy of survivors performed: yes; all animals were necropsied. All animals were euthanized by intraperitoneal injection of sodium pentobarbitone.
- Other examinations performed: clinical signs, body weight
- Mortality/Viability: Four times during test Day 1 (according to the laboratories SOP's the last check was conducted 5 h after application), and daily during Days 2-15.
- Body Weights: Test Days 1 (pre-administration), 8 and 15.
- Clinical Signs: Each animal was examined for changes in appearance and behaviour four times during Day 1, and daily during Days 2-15. All abnormalities were recorded. The animals were checked for the clinical signs.

Statistics:

The LOGIT-Model could not be applied to the observed rates of death. The toxicity was estimated without use of a statistical model.

Preliminary study:

No data

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Remarks on result:

other: other details not available

Mortality:

No premature death occurred.

Clinical signs:

other: Sedation and ruffled fur were observed in all treated animals between 24 h and 4 d after administration of test substance.

Gross pathology:

No macroscopic findings were observed.

Other findings:

No data

None

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 of the test substance was found to be >2000 mg/kg bw in rats.

Executive summary:

A study was conducted to assess the acute oral toxicity of the test substance (of 100 % purity) in HanIbm: WIST (SPF) rats according to OECD Guideline 401and EU Method B.1 in compliance with GLP.

Groups of 10 fasted animals (5/sex/dose) received a single oral (gavage) dose of 2000 mg/kg bw of the test substance. Parameters assessed included mortality, clinical observations, body weight and necropsy findings in all animals after a 15 d observation period.

No mortality occurred and no significant macroscopic abnormalities were seen at necropsy. Further, there were no effect on body weight gain. However, sedation and ruffled fur were observed in all treated animals between 24 h and 4 d after dosing.

Under the study conditions, the oral LD50 of the test substance was found to be >2000 mg/kg bw in rats.

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

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute toxicity – oral

The Key study was conducted to assess the acute oral toxicity of the test substance in Wistar rats according to OECD Guideline 401 and EU Method B.1 in compliance with GLP.

Groups of 10 fasted animals (5/sex/dose) received a single oral (gavage) dose of 2000 mg/kg bw of the test substance. Parameters assessed included mortality, clinical observations, body weight and necropsy findings in all animals after a 15 d observation period.

No mortality occurred and no significant macroscopic abnormalities were seen at necropsy. Further, there were no effect on body weight gain. However, sedation and ruffled fur were observed in all treated animals between 24 h and 4 d after dosing. Under the study conditions, the oral LD50 of the test substance was found to be > 2000 mg/kg bw in rats.

Couple of more studies conducted to assess the acute oral toxicity of the test substance in rats found the oral LD50 of the test substance to be > 5000 mg/kg bw.

Based on the data of all the three studies it can be concluded that, Reactive Black 039 is considered to have low toxicity by oral route.

Further analysis of our substance does not indicate any harm upon oral exposure and neither the chemical structure of the substance nor any toxicokinetic results raise any concern about the toxic behavior of the substance upon dermal absorption, we will not perform any test on this endpoint in in vitro or in vivo test systems.

Acute toxicity – inhalation
Based on Column 2 of the table given in REACH Annex VIII, 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 very low vapour pressure of the substance, the fact that the substance is imported into the EU in a formulated form as a dust-free powder or as a granulate, the inhalation route of exposure is considered to be unlikely, thus the study on acute inhalation toxicity is being waived.

Acute toxicity – dermal
The substance FAT40171 has been tested for acute oral toxicity and has been found to be not toxic to the animals investigated LD50 (males/females) >2000 mg/kg bw. None of the animals died and none of the animals exhibit any notable signs of toxicity. In addition, in animal tests on skin irritation/corrosion effects the test substance has not been found to cause severe irritating or corrosive effects to the skin or any other effect resulting in a destruction of an intact skin barrier. In addition, old tests on skin irritation performed on abraded skin areas of animals resulting in a partial destruction of the skin barrier, also no signs of toxicity were noted. The substance itself is characterized as a dry powdery substance which is marketed in a dedusted form or in a liquid mixture only. The molecular weight of the substance is 1021.1629 g/mole. From basic research on internal and external barriers of humans it is known, that such molecules are almost not able to permeate the skin resulting in an enhanced bioavailability of the substance applied topically. Experts from the chemical industry, CRO s and regulatory authorities, together with the NC 3Rs, have published a review paper highlighting opportunity to waive requirements for acute toxicity testing of non-pharmaceutical chemicals. The review focuses on acute oral, dermal and inhalation toxicity, skin and eye irritation and skin sensitisation. The paper, which is published as an open access article in Critical Reviews in Toxicology, is intended to provide a focused review for the regulatory community to use when considering the need to generate acute toxicity data. Analysis presented in the paper demonstrates that for pesticide active substances and general chemicals, acute dermal toxicity testing very rarely provides information of value for hazard identification or classification and labelling purposes, when an acute oral study has already been conducted. Only 1 out of 438 chemical substances and 2 out of 240 active pesticide substances have been found to be more severe than after oral application. These findings suggest that acute dermal toxicity studies should not be performed except in exceptional circumstances, for example where information on absorption, toxicokinetics or mode of action suggests that acute toxicity might be greater by the dermal rather than oral route. Since analysis of our substance does not indicate any harm upon oral exposure and neither the chemical structure of the substance nor any toxicokinetic results raise any concern about the toxic behaviour of the substance upon dermal absorption, we will not perform any test on this endpoint in in vitro or in vivo test systems.

Justification for classification or non-classification

Based on the available data, Reactive Black 039 does not warrant classification for acute toxicity as per the criteria of Regulation EC No. 1272/2008 (CLP).