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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
study is not fully compliant with current guidelines.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1967
Report Date:
1967
Reference Type:
other: summary report
Title:
Unnamed
Year:
1967
Report Date:
1967

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Test materials used in this dossier are all considered to fall under the definition of nano-materials according to the European Commission Recommendation 2011/696/EU as the synthesis and manufacturing of this pigment always yields particulate material with a fine particle size distribution.

Test animals

Species:
rat
Strain:
other: albino
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Mean weight at study initiation: 130 g (males) and 121 g (females)
- Housing: Makrolon cages on wood shavings, 5 animals per cage
- Diet (e.g. ad libitum): ALTROMIN R, manufactured by Altromin GmbH in Lage/Lippe
- Water (e.g. ad libitum): tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 - 25
- Humidity (%): 35 - 60
- Air changes (per hr): partially air-conditioned animal rooms

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): pelleted feed was freshly prepared at 7-8-day intervals
- Mixing appropriate amounts with (Type of food): ALTROMIN R, manufactured by Altromin GmbH in Lage/Lippe
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Dose / conc.:
10 000 ppm
Remarks:
1% in feed
Dose / conc.:
50 000 ppm
Remarks:
5% in feed
No. of animals per sex per dose:
15
Control animals:
yes, plain diet

Examinations

Observations and examinations performed and frequency:
Each rat was monitored daily to determine its behaviour and general state of health. The body weight of each animal was determined once a week and the food consumption at 7-day intervals.
Before the start and at the end of the trial haematological and urine investigations were carried out in 10 male and 10 female rats from each group. The haematological investigation included determination of the haemoglobin content, number of red and white blood cells as well as the evaluation of the differential blood count and the possible presence of Heinz bodies. The urine investigation covered appearance, colour, protein and sediment.
Sacrifice and pathology:
At the end of the trial all the animals were killed and autopsied and the organs were subjected to histological examination.
Macro- and microscopic examination of the organs heart, lung, liver, kidney, adrenal glands, spleen, cerebrum, cerebellum, testes and ovaries, pancreas, pituitary, thyroid gland, stomach and small intestine.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
All the animals displayed normal behaviour during the trial and no signs of a toxic effect were observed which would have been caused by the pigment. The pigment was excreted via the faeces.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
There were isolated spontaneous deaths during the trial. One rat (animal no. 304) from group 1 (5%) died after 33 trial days and a further rat (animal no. 337) after 73 trial days. One rat (animal no. 362) from group III (control) died after 23 trial days. The rats died of pneumonia. No connection between the substance administered and the deaths could be detected.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The weight gains of the male and female rats from groups I (5%) and II (1%) were normal and corresponded to those of the control animals.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
The food consumption of the experimental animals was the same as that of the control rats.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
The haematological investigations showed no pathological findings even at the end of the trial.
Clinical biochemistry findings:
not examined
Urinalysis findings:
no effects observed
Description (incidence and severity):
The urine investigations showed no pathological findings at the end of the trial. The male rats showed slight protein excretion at the end of the trial which is a physiological feature of adult male rats.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
The organ weight gains of the male and female rats from groups I (5%) and II (1%) were normal and corresponded to those of the control animals.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Macroscopic examination of the organs of rats that had received the pigment in their feed in a 90-day trial revealed no pathological organ damage. Inflammatory lung changes were detected at autopsy in experimental animals no. 304 and 337. Animal no. 322 displayed a liver change for which no reason could be found. The controls no. 360 and 362 also had inflammatory lung processes arid the liver of no. 400 was interspersed with yellowish spots the size of a pinhead. The striking feature of no. 413 was very marked liver lobule patterning.
Neuropathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Histological examination of the organs of rats that had received the pigment in their feed in a 90-day trial revealed no pathological organ damage. The lymphocytic infiltrates frequently observed in the heart, liver, kidneys and intestine are a manifestation of quiescent infections. The perifollicular hyperplasia in the spleen should also be assessed as a non-specific defense reaction of this type. The histo-lymphocytic granulomas found in the liver may be a manifestation of a pre-existing infection, most probably salmonellosis. The lung changes found are characteristic of rat-specific pneumonia. In many of the animals there was impaired spermiogenesis in the form of varying degrees of desquamation of prespermatid cells. The striking frequency in the control group (11 cases) and in the group that received 5% test material (9 cases) and the only isolated occurrence in the group that was given 1% test item (2 cases) indicates that these changes are not attributable to the test substance. In addition, the changes in the brain in the form of meningoencephalitis occasionally observed developed in some cases with foci of glial proliferation independent of the study. These brain changes are diagnosed in rats more frequently than random findings, without causing clinical symptoms. The thyroid changes in the form of interfollicular solidification, associated with formation of follicles to varying degrees, occurred in both the control and the experimental animals.

In summary, it can be stated that no obvious study-related changes were observed. The test item therefore caused no organ-damaging effects in rats in the dosages given and over the selected period of administration.

Effect levels

Dose descriptor:
NOAEL
Effect level:
50 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects were reported

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
In summary, it can be stated that no obvious study-related changes were observed. The test substance therefore caused no organ-damaging effects in rats in the dosages given and over the selected period of administration.