Registration Dossier

Administrative data

Description of key information

Oral (rat): NOAEL = 551mg/kg/day (male), 630 mg/kg bw/day (female) 90 days, National Toxicology Program 1982
Oral (mouse): NOAEL = 200 mg/kg bw/day, 90 days mouse male/female, National Toxicology Program 1982

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
April 1977 to May 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: A non-GLP study conducted to sound scientific principles with the read across substance, bis(2-ethylhexyl) adipate.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
(environmental paprmeters were outside guideline ranges, the chronic toxicity studies were not long enough in duration, only two dose levels were used in the main study and heamotology and blood parameters were not analysed)
GLP compliance:
no
Limit test:
no
Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: NCI Frederick Cancer Research Center, Frederick, Maryland, USA
- Age at study initiation: 4 weeks
- Housing: 5 animals per cage in solid bottom suspended polycarbonate cages
- Diet: powdered Wayne® Lab Blox diet, ad libitum
- Water: ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 31 °C
- Humidity (%): 10 - 88 %
- Air changes (per hr): 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
Test diets were prepared by mixing the test material with an aliquot of diet, the mixture was then placed into a Patterson-Kelly® twin-shell intennsifier bar V-blender with the remainder of the feed, and mixed for 10 minutes.
- Storage temperature of food: Test diets were dealed in labelled plastic bags and stored at 4 °C for no longer than 14 days.

Stability of test material in feed was determined by analysing sample diet mixtures containing 100,000 ppm test material that had been stored at -20, 5, 25 or 45 °C for 2 weeks. The amounts of test material found to be present by vapour-phase chromatography indicate that the test material was stable in feed for 2 weeks at temperatures as high as 45 °C.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The amounts of test material in selected batches of feed were measured by vapour-phase chromatography. 2 g samples of feed were extracted with 50 mL portions of methanol. The supernatant solutions were combined and diluted to a volume of 100 mL and analysed by vapour-phase chromatography using the following system:
- Instrument: Bendix 2500
- Column: 3 % OV-17 on 80/100 Supelcoport, glass, 1.8 m x 4 mm ID
- Oven temperature: 250 °C
- Retention time of test material: 1.9 minutes
- Inlet temperature: 235 °C
- Detctor temerature: 250 °C
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
0, 1600, 3100, 6300, 12500, 25000 ppm
Basis:
nominal in diet
No. of animals per sex per dose:
10 males and 10 females per dose
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: dose selection for the 13 week repeated dose toxicity study was based on findings of the preceding acute and 14 day repeated dose toxicity studies (see below)

ACUTE TOXICITY STUDY
The estimated LD50 for male and female rats was 15.0 and 24.6 g/kg, respectively.

14 DAY REPEATED DOSE TOXICITY STUDY
All female mice fed 100,000 ppm died. Weight loss occurred among male mice fed 50,000 ppm and females fed 25,000 ppm or more. Feed consumption was reduced in females fed 100,000 ppm..
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No


OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
SACRIFICE
At the end of the 91 day study, survivors were sacrificed. Necropsies were performed on all animals, and tissues were taken for histopathological analysis.

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):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
One female mouse died as a result of an accident. Weight gain depression was 10 % or more for male mice fed 3,100 ppm or more. Weight gain depression was 10 % or more for females fed 6,300 or 25,000 ppm. No treatement-related histopathologic effects or reduction in feed consumption were observed.
Dose descriptor:
NOAEL
Effect level:
1 600 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Decreased body weight gain.
Critical effects observed:
not specified

Table 1: Dosage, Survival, and Mean Body Weight Data

Sex

Males

Females

Dose (ppm)

Survival

Mean body weight (g)

Weight change relative to controls (%)

Survival

Mean body weight (g)

Weight change relative to controls (%)

Initial

Final

Gain

Initial

Final

Gain

0

10/10

20.2

33.3

13.1

10/10

16.8

25.3

8.5

1,600

10/10

20.7

33.5

13.5

3.1

10/10

16.8

25.0

8.2

-3.5

3,100

10/10

20.7

30.7

10.0

-24

10/10

16.8

25.6

8.8

3.5

6,300

10/10

20.7

32.4

11.7

-10.7

10/10

16.8

21.5

4.7

-45

12,500

10/10

20.7

31.8

11.1

-15.3

9/10 *

16.8

25.8

9.0

5.6

25,000

10/10

20.7

30.5

9.8

-25.2

10/10

16.8

24.2

7.4

-13

* accidental death

Conclusions:
Under the conditions of the study, the No Observed Adverse Effect Level was determined to be 1,600 ppm, based on decreased body weight gain, which is equivalent to approximately 200 mg/kg bw/day for male and female mice.
Executive summary:

The repeated dose toxixcity of the test material was determined in a non-GLP study. During the test, groups of 10 male and 10 female mice were administered with test material in the diet at dose levels of 0 (plain diet), 1,600, 3,100, 6,300, 12,500 and 25,000 ppm, for a period of 13 weeks. Clinical observations were performed twice daily and body weights were recorded weekly. At the end of the 91 day study, survivors were sacrificed. Necropsies were performed on all animals, and tissues were taken for histopathological analysis.

One female mouse died as a result of an accident. Weight gain depression was 10 % or more for male mice fed 3,100 ppm or more. Weight gain depression was 10 % or more for females fed 6,300 or 25,000 ppm. No treatement-related histopathologic effects or reduction in feed consumption were observed. Under the conditions of the study, the No Observed Adverse Effect Level was determined to be 1,600 ppm, based on decreased body weight gain, which is equivalent to approximately 200 mg/kg bw/day for male and female mice.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subchronic
Species:
mouse
Quality of whole database:
The study was not performed to GLP but was conducted to sound scientific principles with a good level of reporting. Since there were a number of deviations from standardised guidelines, and as the study was conducted with the read across substance, bis(2-ethylhexyl) adipate, the study was assigned a reliability score of 2 according to the criteria of Klimisch.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The repeated dose toxixcity of the test material was determined in a non-GLP study. During the test, groups of 10 rats of each sex, and groups of 10 mice of each sex, were administered with test material in the diet at dose levels of 0 (plain diet), 1,600, 3,100, 6,300, 12,500 and 25,000 ppm, for a period of 13 weeks. Clinical observations were performed twice daily and body weights were recorded weekly. At the end of the 91 day study, survivors were sacrificed. Necropsies were performed on all animals, and tissues were taken for histopathological analysis.

 

In rats, one female receiving 1,600 ppm died, but its death was not considered to be treatment-related. Weight gain depression was 11 % or more for male rats fed 12,500 or 25,000 ppm. No test material-related histopathological effects or reduction in feed consumption were observed. Under the conditions of the study, the No Observed Adverse Effect Level was determined to be 6300 ppm which is equivalent to 551 mg/kg bw/day for female and 630 mg/kg bw/day for male rats, respectively.

 

In mice, one female died as a result of an accident. Weight gain depression was 10 % or more for male mice fed 3,100 ppm or more. Weight gain depression was 10 % or more for females fed 6,300 or 25,000 ppm. No treatement-related histopathologic effects or reduction in feed consumption were observed. Under the conditions of the study, the No Observed Adverse Effect Level was determined to be 1,600 ppm, based on decreased body weight gain, which is equivalent to approximately 200 mg/kg bw/day for male and female mice.

In accordance with Section 1 of Annex XI further repeated dose toxicity studies, as required under Section 8.6.2 of Annex IX do not appear scientifically necessary. The existing oral data is considered to adequately address the repeated dose toxicity endpoint.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The study was selected as the source of the key value, since the mouse was the most sensitive of the species tested.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the test material does not require classification for specific organ toxicity, repeated dose. The effects observed in the main study are not considered to be toxicologically significant and do not indicate any signs of organ dysfunction.