PIGMENTADDITIV RL

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 30 Mar to 27 Apr 1989

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD TG 407), according to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

(adopted 1981)

GLP compliance:

yes

Limit test:

no

Test material

Reference

Name:

Unnamed

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: HOECHST AG, Kastengrund, Germany
- Age at study initiation: approx. 6 weeks
- Weight at study initiation: males: mean 123 - 125 g; females: mean 115 - 122
- Fasting period before study: no
- Housing: groups of 5 per sex
- Diet (ad libitum): Altromin 1324 rat diet (Altromin GmbH, Lage/Lippe, Germany)
- Water (ad libitum): tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 50 +/- 20
- Air changes (per hr): not stated, fully air-conditioned
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 2% starch mucilage

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: daily

VEHICLE
- Justification for use and choice of vehicle (if other than water): test item not soluble in water
- Concentration in vehicle: 0, 1.25, 5, 20 % (w/v)
- Amount of vehicle (if gavage): 5 mL/Kg bw

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

28 days

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 62.5, 250, 1000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: after single application of 2000 mg/kg bw of the test item to rats no animal died within the 14 d observation period and no clinical signs of toxicity were observed; based on these findings the doses for the subacute toxicity study were selected

Positive control:

none

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: behaviour and general health status at least daily (twice daily during work days and once daily during weekend and at holidays); weekly check neurological disturbances, eye opacity, damage of oral mucosa and impairment of dental growth
- Cage side observations not reported in detail.

DETAILED CLINICAL OBSERVATIONS: No


BODY WEIGHT: Yes
- Time schedule for examinations: before treatment and 2 times per week during exposure period

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes (measurement 2 times per week)
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: weekly; water consumption over 16 h was determined weekly

OPHTHALMOSCOPIC EXAMINATION: Yes (turbidity)
- Time schedule for examinations: weekly
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: before final sacrifice
- Anaesthetic used for blood collection: Yes (nembutal)
- Animals fasted: No
- How many animals: all
- Parameters examined: erythrocytes count, haemoglobin, haematocrit, leucocytes count, thrombocytes count, differential blood count, reticulocytes (only control and high dose group), Heinz bodies (only control and high dose group), mean clotting time, MCV, MCH, MCHC

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: before final sacrifice
- Animals fasted: No
- How many animals: all
- Parameters examined: sodium, potassium, inorganic phosphorus, uric acid, total bilirubin, creatinine, glucose, urea, calcium, chloride, GOT, GPT, alkaline phosphatase, gamma glutamyl transpeptidase, total protein, albumin and globulin

URINALYSIS: Yes
- Time schedule for collection of urine: overnight from day 26 to 27 of the study
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined: appearance, colour, pH, hemoglobin, protein, glucose, ketone bodies, bilirubin, urobilinogen, specific gravity, sediment (only control and high dose group)

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
- organ weights: heart, lung, liver, kidney, spleen, testes, adrenals

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (heart, lung, liver, kidney, spleen, stomach, jejunum, colon, thymus, testes, adrenals, bone marrow (femur))

Statistics:

Body weights at the designated measurement times, haematological data, clinical chemistry parameters, absolute organ weights and organ to body weight ratios, pH and specific gravity of urine were evaluated with the aid of a pro- gram package for the evaluation of toxicological studies, according to the respective standard operating procedures.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
- no adverse effects
- faeces was discoloured blue from day 6 of the experiment in the mid and high dose group
- no compound-related neurological disturbances, opacity of the refracting media of the eyes, impairment of dental growth or changes of the oral mucosa were observed


BODY WEIGHT AND WEIGHT GAIN
- no effects

FOOD CONSUMPTION
- no effects

FOOD EFFICIENCY
- not examined

WATER CONSUMPTION
- no effects

OPHTHALMOSCOPIC EXAMINATION
- no effects

HAEMATOLOGY
- no adverse effects
- mean clotting time was statistically significant increased in males of the mid dose group, but all values were within the range of the historical controls and no such effect observed in other dose groups or in females

CLINICAL CHEMISTRY-
- no adverse effects
- statistically significant decrease of bilirubin concentration in females of high dose group; this effect was not regarded to be meaningful as only an increase would be toxicologically relevant
- statistically significant increased alpha-1 globulin and decreased albumin and albumin/globulin ratio in males of the high dose group; these effects were not regarded to be relevant by the authors of the study as there were no correlating histopathological or clinical findings

URINALYSIS
- no effects

NEUROBEHAVIOUR
- not examined

ORGAN WEIGHTS
- no adverse effects
- statistically signifcant increased relative (but not absolute) lung weights in males of the high dose group (0.475 +/- 0.062; 0.535 +/- 0.069; 0.466 +/- 0.042; 0.597 +/- 0.096 % body weight in the control, low, mid, high dose group respectively); no such effect occurred in females, no histopathological findings, therefore the effects were not regarded to be of toxicological relevance

GROSS PATHOLOGY
-no effects

HISTOPATHOLOGY: NON-NEOPLASTIC
- no adverse effects
- only incidential observations

Effect levels

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects observed up to the highest dose tested

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Under the conditions of this study the test item did not induce adverse treatment related effects after 28 days of exposure to doses up to 1000 mg/kg bw/d.

Executive summary:

Groups of 5 male and 5 female Wistar rats received the test item in a subacute study according to OECD guideline 407 and GLP by oral gavage at dose levels of 0, 62.5, 250 or 1000 mg/kg body weight per day for 28 days and were necropsied at day 29. There were no clinical signs or mortality. Blue discolouring of the faeces occurred in males and females of the mid and high dose group starting on day six of treatment. Behaviour, health status, body weight development, food and water consumption were not impaired in males and females of all groups. Clinical chemistry revealed a statistically significant decrease of bilirubin concentration in females of high dose group. This effect was not regarded to be meaningful as only an increase would be toxicologically relevant. A statistically significant increase of alpha-1 globulin and a decrease of albumin and albumin/globulin ratio was observed in males of the high dose group. These effects were not regarded to be relevant as there were no correlating histopathological or clinical findings. The haematological examinations did not reveal any substance related changes except statistically significant increase of the mean clotting time in males of the mid dose group. As all values were within the range of the historical controls and no such effect was observed in other dose groups or in females they were not regarded to be of any relevance. No treatment-related changes were detected by urinalysis. The administration of the test compound did not affect absolute or relative organ weights except of the lung weights in males. The absolute and and relative lung weights were increased in males of the high dose group. But only the effect on relative lung weights revealed statistical significance. As there were no histopathological correlations and no such effects occurred in female rats the effects were judged not to be of toxicological relevance. No compound-related macroscopically visible changes were found at necropsy. The histopathological examinations also revealed no substance-related changes. Under the conditions of this study the test item did not produce adverse treatment related effects after 28 days of exposure to doses up to 1000 mg/kg bw/day.