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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
GLP compliance:
yes (incl. certificate)
Remarks:
testing lab.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Trilon C Flussig
Containing 43.7% "active", pentasodium DTPA

tank 64, from October 26, 1989

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Male and female Wistar rats (Chbb:THOM (SPF)) supplied by Dr . Karl Thomae GmbH, Biberach/Riss, Germany, which were free of signs of disease were used for the investigations.
The rats were identified clearly by tattooing of the respective animal number into the ears.
The rats were housed singly in type DK III stainless steel wire mesh cages supplied by Becker & Co., Castrop-Rauxel, Germany (floor area about 800 cm2). Underneath the cages, waste trays were fixed containing absorbent material (type 3/ dustfree embedding, supplied by SSNIFF, Soest, Germany). The animals were housed in a fully air-conditioned room. Central air-conditioning guaranteed a range of 20 - 24°C for temperature and of 30 - 70% for relative humidity. The day/night rhythm was 12 hours (12 hours light from 06.00 a.m. - 06.00 p.m., 12 hours dark from 06.00 p.m. - 06.00 a.m.). Deviations from these ranges did not occur. The animal room was completely disinfected using a disinfector ("AUTEX", fully automatic, formalin-ammoniabased
terminal disinfector). The floor and the walls were cleaned once a week. The cleansing liquid used was water containing about 0.1 % Incidin perfect® (supplied by Henkel, Düsseldorf, Germany). The food used was basic maintenance diet rat/mouse/hamster, meal, supplied by Klingentalmühle AG, Kaiseraugst, Switzerland. Food and drinking water (from water bottles) were available ad libitum.

Administration / exposure

Route of administration:
oral: drinking water
Details on oral exposure:
The test substance was administered as a solution in drinking water.
The test substance was weighed for the specific test group, and the appropriate amount of drinking water was added (also weighed). This mixture was subsequently stirred with a magnetic stirrer for at least 30 minutes to reach complete solubility of the test substance in the drinking water. The drinking water solutions were prepared twice a week (Monday to Friday).
Before the beginning of the study, the stability of the test substance in the vehicle over a period of 4 days at room temperature was ordered. At the start of the study one sample of each concentration was sent to the analytical laboratory for determination of the correctness of the concentration of the test substance preparations.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability of the test substance in the vehicle over a period of 4 days at room temperature and the correctness of the concentrations were verified analytically.
Duration of treatment / exposure:
4 weeks
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
600 ppm; 3,000 ppm and 12,000 ppm
Basis:

No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
Selction of the doses for this study was based on the results of a previous palatability study.
In this study, Trilon C flüssig was administered in the drinking water to 3 male and 3 female Wistar rats per group for 2 weeks .
Following substance-induced changes were observed:
15,000 ppm:
• Discoloration of the feces in both sexes
• Decreased food consumption in both sexes
• Decreased water consumption in both sexes
• Piloerection and reduced general state in 1 male and 1 female
• Diminished adipose tissue in 1 male and 2 females
5,000 ppm:
• Discoloration of the feces in both sexes
Therefore, the following doses were chosen for the 4-week administration in the drinking water (doses of active ingredient):
600 ppm: as the lowest dose level
3,000 ppm: as the intermediate dose level
12,000 ppm: as the high dose, resulting in a substance intake of > 1,000 mg/kg body weight
Positive control:
none

Examinations

Observations and examinations performed and frequency:
Clinical observations: A check was made twice Mondays to Fridays and once a day on Saturdays, Sundays and public holidays for general observations. Once a week an additional comprehensive clinical examination was carried out. In order to prepare tables of the clinical symptoms observed, the data were entered offline into the computer systems; according to the particular health status of the individual animals, sometimes several, different clinical signs were documented during one week.
Mortality: A check was made twice Mondays to Fridays and once a day on Saturdays, Sundays and public holidays for any dead or moribund animals.
CLINICAL PATHOLOGY: Blood was taken from the retroorbital venous plexus in the morning from fasted animals without anesthesia. The blood sampling procedure and the subsequent analysis of the blood and serum samples were carried out in a randomized sequence.
The following parameters were determined in blood with EDTA-K3 as anticoagulant using a particle counter (S Plus model, by Coulter, Krefeld, Germany) :
• leukocytes
• erythrocytes
• hemoglobin
• hematocrit
• mean corpuscular volume
• mean corpuscular hemoglobi n
• mean corpuscular hemoglobin concentration
• platelets
Clinical chemistry: The following parameters were determined :
• alanine aminotransferase
• aspartate aminotransferase
• alkaline phosphatas e
• serum-y-glutamyltransferase
• sodium
• potassium
• chlorid e
• inorganic phosphate
• calcium
• urea
• creatinine
• glucose
• total bilirubin
• total protein
• albumin
• globulins
• triglycerides
• cholesterol
• magnesium
Urinalyses: The following examinations were carried out:
• volume
• color
• turbidity
• nitrite
• pH
• protein
• glucose
• ketones
• urobilinogen
• bilirubi n
• blood
• specific gravity
• sediment
Sacrifice and pathology:
A check was made twice Mondays to Fridays and once a day on Saturdays, Sundays and public holidays for any dead or moribund animals.
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Statistics:
Statistics for clinical examinations, clinical chemistry and hematology, urinalyses.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
no effects observed
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:
effects observed, treatment-related
Urinalysis findings:
effects observed, treatment-related
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
12,000 ppm (about 1,775 mg/kg body weight): discoloration of the feces in both sexes; decreased food consumption in both sexes; significantly decreased body weight, resulting in reduced values of about 22% (males) and 7% (females) at the end of the study; significantly reduced body weight change, resulting in reduced values of about 46% (males) and 21 % (females) at the end of the study; increase in alanine aminotransferase in the males; increase in specific gravity, renal epithelial cells and casts in the urines of individual males; dark yellow colored urines in individual males; decrease in urine volume in both sexes; decrease in alkaline phosphatase in the females; transitional cell hyperplasia in the urinary bladder of 4 male and 2 female test animals.
3,000 ppm (about 420 mg/kg body weight): significantly decreased body weight change in the males in the last test week (about 10% below controls); increase in alanine aminotransferase in the males; decrease in alkaline phosphatase in the females.
600 ppm (about 75 mg/kg body weight): no substance-induced changes were detected.

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
ca. 75 mg/kg bw (total dose)
Sex:
male/female
Basis for effect level:
other: Clear changes in body weight, clinical chemistry, urinalaysis, and histopathology in the high dose group. Clear changes in body weight and clinical chemistry in the mid dose group.

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Clear effects of body weight and histopathological changes of the urinary tract with corroborating results of the urinalyses were obtained at 12,000 ppm. At 3,000 ppm, only minor effects were obtained. Although the clinicochemical findings at 12,000 and 3,000 ppm are difficult to explain, they must be assessed as substance-related. A clear "no observed adverse effect level" was obtained at 600 ppm (about 75 mg/kg body weight) under the test conditions chosen.