Anthraquinone

Endpoint summary

Description of key information

One 90 days study of repeated dose toxicity by oral route was performed in 2018. The LOAEL of 50 mg/kg bw/day was established.

There are two other studies of repeated dose toxicity by oral route for Anthraquinone available (as study summary) with these results:
NOAEL: 2 mg/kg bw/day (28-day study)
NOAEL: 1.36 mg/kg bw/day (90-day study)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1975-6

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: only study summary available and IUCLID data set summary available

Qualifier:

no guideline available

Principles of method if other than guideline:

no data

GLP compliance:

not specified

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

other: water and Cremophor

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
2, 10, 20, 50, 250 mg/kg
Basis:
actual ingested

No. of animals per sex per dose:

15 females and 15 males

Control animals:

yes, concurrent vehicle

Details on study design:

Two experiments were performed. The dosage in first experiment was 10, 50, 250 mg/kg. The dosage in second experiment was 2 and 20 mg/kg. The application volume was 10 mL/ kg body weight.

Observations and examinations performed and frequency:

CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes

HAEMATOLOGY: Yes

CLINICAL CHEMISTRY: Yes

URINALYSIS: Yes

Details on results:

RESULTS
All dose groups: no deaths
2 mg/kg bw/day: no signs of toxicity
10, 20, 50, and 250 mg/kg bw/day: impairment of the general condition, black-colored spleen, splenic congestion, increased relative weights of the liver and the spleen
10 mg/kg bw/day: hepatocyte enlargement
10, 50, and 250 mg/kg bw/day: increased relative renal weights in the females
20, 50, and 250 mg/kg bw/day: decreased body weight gain in the females, erythropenia
50 and 250 mg/kg bw/day: decreased body weight gain in the males, increased relative weights of the thyroid, the heart, the testes and the kidneys in the males, hepatocyte enlargement
250 mg/kg bw/day: decreased relative weights of the ovaries, clinical chemistry: slightly increased concentrations of glutamate-pyruvate transaminase and of glutamate oxalo-acetate transaminase.

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 2 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

organ weights and organ / body weight ratios

Critical effects observed:

not specified

Conclusions:

After administration of the substance orally, impairment of the general condition, black-colored spleen, splenic congestion, increased relative weights of the liver and the spleen were recorded in the treated animals (except the lowest dose level). These histopathological findings were recorded:
10 mg/kg bw/day: hepatocyte enlargement
10, 50, and 250 mg/kg bw/day: increased relative renal weights in the females
20, 50, and 250 mg/kg bw/day: decreased body weight gain in the females, erythropenia
50 and 250 mg/kg bw/day: decreased body weight gain in the males, increased relative weights of the thyroid, the heart, the testes and the kidneys in the males, hepatocyte enlargement
250 mg/kg bw/day: decreased relative weights of the ovaries, clinical chemistry: slightly increased concentrations of glutamate-pyruvate transaminase and of glutamate oxalo-acetate transaminase.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1978-9

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: only study summary available and IUCLID data set summary available

Qualifier:

no guideline available

Principles of method if other than guideline:

no data

GLP compliance:

not specified

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: feed

Duration of treatment / exposure:

3 months

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
15, 150, 1500 ppm (=ca. 1, 10, 100 mg/ kg bw.)
Basis:
nominal in diet

No. of animals per sex per dose:

20 females and 20 males

Control animals:

yes

Observations and examinations performed and frequency:

CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes

FOOD CONSUMPTION AND COMPOUND INTAKE : Yes

HAEMATOLOGY: Yes

CLINICAL CHEMISTRY: Yes

Details on results:

RESULTS:
All dose groups: no deaths
15 ppm (= 1.36 mg/kg/bw/day): no symptoms of toxicity
150 and 1500 ppm (=about 10 and 100 mg/kg bw/day): decreased food intake, increased absolute weights of the liver
1500 ppm (=about 100 mg/kg bw/day): decreased body weight gain, enlargement of the centrilobular hepatocytes; clinical chemistry: increased cholesterol levels at the end of the test period mainly in the females.

Dose descriptor:

NOAEL

Effect level:

15 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

food consumption and compound intake

organ weights and organ / body weight ratios

Critical effects observed:

not specified

Conclusions:

After application of the substance in diet, decreased body weight gain, decreased food intake, increased cholesterol levels at the end of the test period mainly in the females, increased absolute weights of the liver and enlargement of the centrilobular hepatocytes were recorded.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEL

50 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

chronic toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1970

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.29 (Sub-Chronic Inhalation Toxicity:90-Day Study)

Deviations:

yes

Remarks:

see the description below in the field "Principles of method if other than guideline"

Principles of method if other than guideline:

The rats were treated by dynamic inhalation introduction of Anthraquinone dust into the chamber, designed by I. F. Bojarčuk. The animals were exposed to Anthraquinone during 4 months, 5-6 hours daily. Atmospheric conditions were controlled by chemical method with sensitivity of 5 micrograms in 3 ml of assessed volume. Study was performed with doses 12.2±0.7 mg/m3 (the most frequently occuring exposure in production conditions) and 5.2±0.8 mg/m3.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

not specified

Route of administration:

inhalation: dust

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

The animals were exposed to Anthraquinone during 4 months, 5-6 hours daily.
Study was performed with doses 12.2±0.7 mg/m3 (the most frequently occuring exposure in production conditions) and 5.2±0.8 mg/m3.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Atmospheric conditions were controlled by chemical method with sensitivity of 5 micrograms in 3 ml of assessed volume.

Duration of treatment / exposure:

5-6 h

Frequency of treatment:

daily, 4 months in total

Dose / conc.:

12.2 mg/m³ air (analytical)

Remarks:

Doses / Concentrations:
12.2±0.7 mg/m³
Basis:
analytical conc.

Dose / conc.:

5.2 mg/m³ air (analytical)

Remarks:

Doses / Concentrations:
5.2±0.8 mg/m³
Basis:
analytical conc.

No. of animals per sex per dose:

total - 96 animals including control group

Control animals:

yes

Details on study design:

Functional state of the nervous system of animals was evaluated according to the general threshold indicator (S. V. Spěranskij): protein spectrum of blood serum, thymol test (Mac Lagan), the level of glycogen in the liver (Formol and Telete) provides information on the functional state of liver, the level of creatinine and residual nitrogen in blood indicated the functional state of kidneys. In addition, clinical blood tests were carried out with respect to regeneration (reticulogram) and degeneration (Heinz bodies, methemoglobin) in components of red blood cells and phagocytic activity of leukocytes. Irritation effect of substance on skin and mucous membranes of the eyes was also evaluated, and photodynamic effect was evaluated in a special series of experiments. Organs of killed animals were subject to histological analysis.

Observations and examinations performed and frequency:

Anthraquinone at concentration of 12.2 mg/m3 had some effects on experimental animals: body weight loss, the level of hemoglobin was decreased to 76.2 and 74.4 % (97% in control group, P <0.05) at the end of 2nd and 4th month of exposure, decrease in the number of eryrthrocytes to 7.5 million (9.08 control trial, P <0.05) and relative reticulopenia. During the first 3 months of treatment the deficiency of vitamin C levels in blood in exposed animals was noted: basic level - 1 mg %, 1st month - 0.6 mg % (P <0.05), 2nd month - 0.45 mg %, 3rd month - 0, 39 mg %, in control experiment - 1 ÷ 0.8 mg %.
Concentration 5.2 ± 0.8 mg / m³: No significant changes.

Sacrifice and pathology:

Concentration 12.2 ± 0.7 mg / m³:
During histological analysis in the lungs of animals killed after treatment emphysema and atelectasis, cellular proliferation, in particular perivascular hyperemia of the capillaries and exsudation in the alveolar lumen. Blood picture was normalized during the experimental period, changes of the lung regenerated within the first month after termination of the experiment.

Concentration 5.2 ± 0.8 mg / m³:
No toxic effects of Anthraquinone were observed; Anthraquinone was regarded as ineffective in chronic experiment.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Haematological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Key result

Dose descriptor:

NOEC

Effect level:

ca. 5.2 mg/m³ air (analytical)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: no changes at this concentration

Dose descriptor:

other: Maximal Acceptance Concentration MAC

Effect level:

ca. 10 mg/m³ air

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: predicted value

Critical effects observed:

not specified

Executive summary:

The obtained experimental data allow to classify Anthraquinone as chemical dust of low toxicity, with no specific effect, because disturbance of trophic processes, tendency to reduce levels of hemoglobin and erythrocytes, deficiency of vitamin C in blood can be observed only when large quantities of dust are administered into the body (subacute test). Due to low toxicity of Anthraquinone, the observations in production combined with the experimental data allow to recommend a maximum permissible concentration of substance in the air of working area of 10 mg/m³.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

5.2 mg/m³

Study duration:

subchronic

Additional information

The toxicity of Anthraquinone was investigated in several studies using the oral (drinking water and feeding) and inhalation routes.

In a 28-day repeated dose toxicity study via oral route, the NOAEL value of 2 mg/kg bw/day and the LOAEL value 10 mg/kg bw/day were determined(Bayer AG, 1976, The Draft Assessment Report – Anthraquinone, Belgium, 2006).

In a 90-day repeated dose toxicity study via oral route,the NOAEL value 1.36 mg/kg bw/day and the LOAEL value 12.6 mg/kg bw/day were determined (Bayer AG, 1979, The Draft Assessment Report – Anthraquinone, Belgium, 2006).

In a 4 -month repeated dose toxicity study via inhalation route, the NOAEC was not determined in the study, although the exposure to the concentration of 5.2 mg/m³ did not elicit any adverse effect in animals (Volodchenko , V.A. et al., 1970). Therefore it could be concluded that NOAEC is 5,2 mg/m3.

The LOAEL (Lowest Observable Adverse Effect Level) value for REPEATED DOSE TOXICITY (90 days) in male and female rats for the test item Anthraquinone was established as 50 mg/kg/day.

The value of LOAEL (Lowest Observable Adverse Effect Level) was established on the basis of results of histopathological examination of liver, haematological examination (decreased values of total ertythrocyte, haemoglobin and haematocrit, prolonged of APTT), biochemical examination (decreased activity of AST, decreased values of bile acids, cholinesterase, bilirubin total, urea and increased values of protein total, albumin, cholesterol total, increased concentration of calcium ions) and biometry of organs (increased weight of liver, kidneys and spleen).

Justification for classification or non-classification

Based on the test results and according to the EC criteria for classification and labelling requirements for dangerous substances and mixtures the test substance Anthraquinone does not have to be classified for repeated dose toxicity.