Registration Dossier

Administrative data

Endpoint:
repeated dose toxicity: oral
Remarks:
other: subacute to subchronic
Type of information:
experimental study
Adequacy of study:
other information
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: meets generally accepted scientific standards, well documented study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1959
Report date:
1959

Materials and methods

Principles of method if other than guideline:
BASF test: evaluation of test substance toxicity after repeated administration (up to 90 days by gavage) to test animals
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Palatinol IC (Di-Iso-butylphthalate)
- Analytical purity: technical grade
No additional detail provided

Test animals

Species:
rabbit
Strain:
other: White Vienna
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: not specified; it is stated that the animals had been previously used in experiments not further specified
- Age at study initiation: means of 3.1 kg
No additional details provided

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
- Concentration in vehicle: the concentration of test substance in vehicle was 25% in the 519.6 and 1039.2 mg/kg and 50% in the mg/kg 2078.4 mg/kg bw dose groups
- Amount of vehicle (if gavage): 2.00 ml/kg in the 2074.4 mg/kg and 2.00 ml/kg in the mg/kg 519.6 and 1039.2 mg/kg bw
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days in the low dose, 5 or 11 days in the mid dose, and 4, 5 or 6 days in the high dose groups (60 days of exposure in the low dose group, 5 [for one male and one female] or 11 days of exposure for the mid dose and 4, 5 or 6 days of exposure for the high dose)
Frequency of treatment:
5 days per week
Doses / concentrations
Remarks:
Doses / Concentrations:
519.6, 1039.2 and 2078.4 mg/kg bw (calculated from 0.5, 1.0 and 2.0 ml/kg bw assuming test substance density of 1.0392 g/ml)
Basis:
actual ingested
No. of animals per sex per dose:
3 females in the low dose), 2 males and 1 female in the mid dose and 3 males in the high dose groups
Control animals:
yes, concurrent vehicle
Details on study design:
no data
Positive control:
none; the test substance was evaluated together with 4 additional

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: start and end

FOOD CONSUMPTION
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: each days -14, 8, 16, 19, 26, 40, 53, 67, 79 and 87
- Anaesthetic used for blood collection: no
- Animals fasted: No
- How many animals: all
- Parameters examined: blood count

CLINICAL CHEMISTRY: No

URINALYSIS: Yes
- Time schedule for collection of urine: days 1, 2, 3, 4, 16, 18, 23, 25, 30, 37, 44, 45, 50, 57, 60, 61, 68, 74, 77 and 84
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Other examinations:
Phthalate excretion in 16-24 urin was also evaluated in the 3 animals of the high dose group
Statistics:
None reported

Results and discussion

Results of examinations

Details on results:
CLINICAL SIGNS AND MORTALITY
- All treated animals died in the mid and high dose groups, none in the low dose group
- Apathy and atony were observed in the mid and high dose groups

BODY WEIGHT (Table 1)
body weight reduction was observed in all dose groups (-0.55% in the low dose, -17.3% in the mid dose and -13.3% in the high dose

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
For all dose groups a small reduction in daily food intake was observed.

HAEMATOLOGY
Increased leukocyte and neutrophile counts were generally observed, essentially in the mid and high dose groups

URINALYSIS
Increased protein was generally observed.
3.55 to 16.4% of phthalate was excreted in urine (see Table 2)

GROSS PATHOLOGY
No significant changes compared to the historical controls were observed in the pathological (gross and histo) analysis.

Effect levels

Dose descriptor:
NOAEL
Effect level:
519.6 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1: Body weights

Dosage

(mg/kg bw)

Mean body weights (kg)

Low dose

Mid dose

High dose

At study start

3.03

3.69

2.59

At the end

3.01

3.05

2.25

Table 2: Phthalate excretion in 16 or 24 hours urine

Percentage of phthalate excreted in urine

First animal

Second animal

Third animal

16 hours

24 hours

16 hours

24 hours

16 hours

24 hours

3.85

7.10

3.55

16.40

4.80

8.50

VEHICLE CONTROL

In the negative control animals receiving 2.0 ml/kg olive oil the two animals died, one after 8, the second after 52 applications (the only symptom shown being body weight reduction in 1 animal). The animals receiving 1.0 ml/kg olive oil were killed after 32 or 50 applications (slight body weight reduction in both animals, sniffing in one animal, strong anemia in both animals). Pneumonia and liver necrosis were observed in 3/4 animals of the control groups at necropsy.

Applicant's summary and conclusion