Registration Dossier

Administrative data

Description of key information

- oral: LD50: > 5000 mg/kg bw (rat; Ciba Geigy Ltd. 1986, BASF AG 1979 and 1980; Lu, 1964; Putulina, 1966)
- inhalation: LC50 > 5.5 mg/L (rat; BASF AG 1990) - 1 of 10 animals died during exposure.
- dermal: LD50 > 1050 mg/kg bw (rat; BASF AG 1975) (testing was performed with a formulation containing 42% of the substance)

In accordance with EC regulation 1907/2006, a substance can be assumed as non-hazardous for the dermal route, if no adverse effects were observed for the oral route at the limit dose. Therefore, for the purpose of risk assessment, the LD50 for the dermal route is considered to be greater than 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
5 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating conc.
5 500 mg/m³

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
1 050 mg/kg bw

Additional information

Oral:

In an acute oral toxicity limit test according to OECD 401 (Ciba Geigy Ltd. 1986; reliability score 2) a group of Sprague-Dawley rats (5 / sex) was given a single oral dose (via gavage) of the test substance in distilled water containing 0.5% carboxymethylcellulose and 0.1% polysorbate at a dose of 5000 mg/kg bw and observed for 14 days. The purity of the test substance was unknown.

No deaths occurred. Moderate to slight dyspnoea, slight exophthalmoses, ruffled fur (slight to moderate), and curved body position were seen, beginning as early as 1 hour post dosing and lasting up to 9 day. The animals were fully recovered within 10 days. The animals gained weight and no deviations from normal morphology were found.

Additionally, there are two studies of BASF AG (1979 and 1980, both: reliability score 2) which were conducted similar to OECD 401.

In the first one groups of Sprague-Dawley rats (5 / sex) were given a single oral dose (via gavage) of the test substance in water at doses of 2150 and 5000 mg/kg bw and observed for 14 days. The test substance was a formulation containing more than 80% of the pigment. The faeces were coloured blue in all doses. Body weight was reduced in female animals of the 2150 mg/kg bw dose only between day 7 and 13 after administration. In all other groups, animals gained weight normally. No abnormalities were detected at pathology and the LD50 was defined as > 5000 mg/kg bw.

In the second BASF test a group of Sprague-Dawley rats (5 / sex) was given a single oral dose (via gavage) of the test substance in distilled water containing 0.5% carboxymethylcellulose at a dose of 5000 mg/kg bw (limit test) and observed for 14 days. The test substance was a formulation containing more than 80% of the pigment. The faeces were coloured blue 24 hours after application of the test substance. This effect was no longer evident from 48 hours till end of the observation period. The animals were all in a good general condition. After 7 days, slight reduction in body weights of male animals was seen. By the next observation time point, body weight gain was normal. No abnormalities were detected at pathology and the LD50 was also defined as > 5000 mg/kg bw. A preliminary test was performed with 1000 and ca. 5000 mg/kg bw using 2 female rats/dose. None of the animals died. No clinical sign of toxicity was reported. No abnormality was seen at gross necropsy.

There are additional data with rats and mice which are not assignable due to insufficiency in documentation, but confirmed the practically nontoxic potential of the test substance (Lu, 1964; Putulina, 1966).

In the RTCES update of 1995 a LD50of 2000 mg/kg bw is described, but this refers to a secondary source (WHO Food additives series) which again refers to the publication of Lu et al, 1964. As described above, Lu et al did not report adverse effects of Pigment Blue 60. Therefore the RTECS data is disregarded.

 

Inhalation:

In an acute inhalation toxicity study (BASF AG 1990; reliability score 1) according to OECD 403 (limit test), a group of male and female Wistar rats (5 animals / sex) were exposed (nose / head only) to a dust aerosol of the test substance at an analytical concentration of 5.5 mg/L for 4 h. The mass median aerodynamic diameter 50% was 1.9 µm, the respirable dust aerosol fraction (reaching the alveolar region) 89%. Animals then were observed for 14 days. Three and a half hours after beginning of exposure, 1 of 5 female animals was found dead. No mortality occurred in the male animals. Clinical signs during exposure were accelerated respiration, irregular respiration, intermittent respiration, eyelid closure, and attempts to escape. Body weight gain over the observation period was comparable to that of historical (air) controls. On day 7 post treatment, body weights were increased by 9.5 and 10% (compared to initial body weights) in female and male animals, respectively) versus 10 and 15% in historical controls. On day 14 post treatment, body weights were increased by 12 and 21% (compared to initial body weights, respectively) versus 19 and 28% in historical controls. Pathological findings in the animal that died were general congestion, atelectasis and cardiac lobe in the lung. No pathological findings were noted at the pathological examinations of the sacrificed animals.

Additionally, there is an inhalation hazard test (IHT) of BASF AG (No. 77/626, 1980; reliability score 2) which was performed in principle as described in OECD test guideline 403 (adopted may 1981). Six male and 6 female rats were exposed to a test atmosphere saturated with dust aerosols of the test substance for 7 hours. The test atmosphere was generated by bubbling 200 L/h dry air through a substance column of about 5 cm above a fritted glass disc in a glass cylinder. The temperature in the exposure chamber was 20°C. The nominal concentration was calculated as quotient of the amount of test substance weight loss during exposure, and the amount of air used during exposure. The concentration of the test atmosphere was stated to be 8.07 mg/L, an analytical verification was not performed. A formulation containing approx. 87% of the test substance was applied. The animals were then observed daily for 7 days for clinical signs and mortality. Weighing was performed at the beginning and end of the study and the animals gained body weight. No mortality resulted from the exposure to the test atmosphere. No clinical signs and no abnormalities at necropsy were observed.

Dermal:

In an acute dermal toxicity study performed in principle similar to OECD 402 (BASF AG 1975; reliability score 2) groups of Sprague Dawly rats (5/sex) were dermally exposed to the test substance in water (a formulation containing < 80% of the test substance was applied) for 24 h at a dose of 1050 mg/kg (active ingredient). An occlusive dressing was used. Animals then were observed for 14 days. Upon removal of the test patches, light blue substance rests were present on the application site. This was no longer seen 4 days later. No mortality and systemic toxicity was observed at this dose.

Additionally, an acute intraperitoneal toxicity study was performed with a formulation containing 87% of the test substance (BASF AG 1980; reliability score 2). 5 mice per sex per dose were administered a single dose (700 and 2000 mg/kg bw) of the test substance formulation via intraperitoneal injection. Clinical signs and body weight are monitored and necropsy is carried out at termination (after 14 days) or after death if an animal died during the study. No mortality occurred. Dyspnoea, apathy, spastic gait, piloerection, convulsions, and poor general state were seen in both dose groups. These symptoms were evident shortly after injection and persisted up to 24 hours after application. The effects were fully reversed 48 hours after application. The sacrificed animals showed intraabdominal test substance incorporations in the liver and slightly coloured adipose tissue.

Taken together in a weight-of-evidence approach, the results of the acute dermal toxicity study with a formulation containing 42% of the test substance and of the acute intraperitoneal toxicity study with a formulation containing 87% of the test substance it is concluded that there is no hazard via the dermal route with a discriminating dose of 2000 mg/kg bw.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC):

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for acute toxicity under Directive 67/548/EEC.

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008:

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. None of the animals died as a result of treatment with the limit dose in the acute oral toxicity study. One of 10 animals died as a result of dust inhalation exposure at the limit dose. No animal died at the highest tested dose tested in the acute dermal toxicity study. As a result the substance is not considered to be classified for acute toxicity under Regulation (EC) No. 1272/2008 as amended for the seventh time in Regulation (EC) No 2015/1221.