Reaction mass of ethoxylated N-oleyl-1,3-propanediamine, hydrofluorides of and ethoxylated N-palmityl-1,3-propanediamine, hydrofluorides of and ethoxylated N-stearyl-1,3-propanediamine, hydrofluorides of and olaflur

Repeated dose toxicity: oral

Administrative data

Endpoint:

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1975

Report date:

1975

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

GLP compliance:

no

Limit test:

no

Test material

Test material information

Test animals

Species:

dog

Strain:

Beagle

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS

- Source: Marshall Research Animals, Inc. North Rose, New York
- Age at study initiation: 5-6 months
- Weight at study initiation: males 8.8-11.8 kg, females 7.4-12.1 kg
- Housing: individually in elevated cages
- Diet (e.g. ad libitum): standard laboratory diet presented daily
- Water: ad libitum
- other: animals were immunized against distemper, hepatitis and leptospirosis (pretest by supplier)

Administration / exposure

Route of administration:

other: oral intubation

Vehicle:

other: 0.25 % Methocel 90 (R) HG Premium 15000 CPS (DOW Chemical)

Details on oral exposure:

Preparation:
Appropriate amounts of compound (adjusted by most recent weekly body weight) were suspended in the vehicle weekly. The suspension was shaken thoroughly before each use.
Storage:
Suspensions were refrigerated when not in use.
Volume:
1 cm³/kg bw days 1-6 ; 3 cm³/kg bw day 7-termination

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

740-749 days

Frequency of treatment:

- daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

1 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

5 mg/kg bw/day (actual dose received)

Doses / concentrations 4

Dose / conc.:

10 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

- 6

Control animals:

yes

Details on study design:

For the first five weeks , the highest dose was 25.0 mg/kg bw/d.
During the 6th week, the high-dose dogs were not dosed to allow them to recover from emesis, diarrhoea and anorexia. During the 7th week, the high dose dogs received placebo treatment (vehicle) in an attempt to decondition dogs from salivating in anticipation of dosing. Dosing was assumed at the beginning of the 8th week at the reduced dose level of 10 mg/kg bw/d.

Examinations

Observations and examinations performed and frequency:

General:
- daily for physical appearance, signs of local systemic toxicity, pharmacologic effects or mortality
Body weight:
- once pretest, weekly and terminally (after fasting)
Food consumption:
- estimated pretest and 4 times weekly thereafter
Ophtalmoscopic examinations:
- pretest and months 6, 12 and 24

Laboratory studies:

- twice pretest and months 1, 3, 6, 9, 12, 20 and 24 (all animals)
Hematology
-hemoglobin, hematocrit, erythrocytes, reticulocytes, total and differential leukocytes, erythrocyte morphology, bone marrow differential, prothrombin time
Clinical chemistry:
- SGOT, SGPT, alkaline phosphatase, blood urea nitrogen, fasting glucose, cholesterol (months 12+24), triglycerides (months 12+24), total protein, creatinine (months 12+24), uric acid (months 12+24), sodium, potassium, calcium, phosphorus, chloride
Urinalysis:
- bilirubin, glucose, gross appearance, ketones, occult blood, pH, protein, specific gravity

Sacrifice and pathology:

- Animals Dying spontaneously: necropsy performed
- Interim necropsy examination: month 6 , No of animals 16 (2/sex/group)

- Sacrifice method: exsanguinated under sodium pentobarbital anaesthesia
- Organs weighed: liver, kidneys, adrenals, thyroid
- Tissues fixed: adrenal, bone rib junction, mandible with teeth, head of femur, bone marrow, sternal, brain, eyes with optic nerve, gall bladder, heart with coronary vessels, colon, duodenum, ileum, jejunum, kidney, liver lungs, lymph node (mesenteric), mammary gland, ovaries, pancreas, parathyroid, pituitary, prostate, salivary gland, skeletal muscle, spleen, stomach, testes, thyroid, urinary bladder, uterus, gross lesions, tissue masses

Statistics:

Body weight, food consumptions, haematology and clinical chemistry parameters, organ weights and organ/body weight ratios were analyzed.
Compound treated groups were compared to control at each time interval.

Results and discussion

Results of examinations

Details on results:

At the original high-dose level (25.0 mg/kg bw/d for the first 5 weeks of test), a decrease in body weight and food consumption occurred, with excessive salivation before and after dosing and increased incidences of diarrhoea and emesis in both sexes were noted. These effects gradually disappeared after the high-dose was reduced from 25.0 to 10 mg/kg bw/d.
After 24 months slightly lower serum phosphorus levels were noted at 10 mg/kg bw/d in both sexes and at 1.0 and 5.0 mg/kg bw/d in the females. Lower phosphorus levels were also noted in the high-dose females at twenty months.
Yellow discoloration and reticoloendothelial cell hyperplasia and hypertrophy of mesenteric lymph nodes occurred in dogs receiving 5.0 and 10.0 mg/kg bw/d.
There were no other microscopic or macroscopic tissue alterations considered attributable to the administration of amine fluorides 297/242.

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

1 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

2

Dose descriptor:

LOAEL

Effect level:

5 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

After 5 weeks, the high dose (25 mg/kg bw/day) was reduced to 10 mg/kg bw/day because of intolerance.

Applicant's summary and conclusion

Executive summary:

In a chronic toxicity study Amine fluorides (1.315 % Olaflur as leading substance and 0.347 % Hetaflur) were administered orally per intubation to 6 Beagle dogs per sex per dose at dose levels of 0, 1, 5 or 25 (10) mg/kg bw/day for a period of 2 years (Menley & James Laboratories, 1975). The NOEL was considered to be 1 mg/kg bw/day for both males and females.

After 5 weeks, the high dose (25 mg/kg bw/day) was reduced to 10 mg/kg bw/day because of intolerance. Animals of the high and middle dose group showed reduced body weight gains. At the end of treatment slightly lower serum phosphate levels were determined as compared to the controls. On dissection, a yellowish discoloration and hyperplasia/hypertrophy of the reticulo-endothelial system was seen. No other macroscopic or microscopic changes were found in the organs and tissues.