2-Butenedioic acid (E)-, di-C8-18-alkyl esters

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

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 1

Reference Type:

study report

Title:

Unnamed

Year:

1982

Reference 2

Reference Type:

secondary source

Title:

Diesters Category of the Aliphatic Esters Chemicals (Test Plan and Robust Summaries for Substances in the HPV Test Plan)

Author:

US-EPA (American Chemistry Council's Aliphatic Esters Panel)

Year:

2010

Bibliographic source:

High Production Volume (HPV) Chemical Challenge Program (201-16837A and 201-16837B)

Reference 3

Reference Type:

secondary source

Title:

Bis(2-ethylhexyl)adipate (DEHA) CAS N°: 103-23-1

Author:

OECD

Year:

2000

Bibliographic source:

SIDS Initial Assessment Report for SIAM 10; Tokyo, Japan, 15-17 March 2000

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

no data on haematology and clinical chemistry

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Bis(2-ethylhexyl) adipate

EC Number:

203-090-1

EC Name:

Bis(2-ethylhexyl) adipate

Cas Number:

103-23-1

Molecular formula:

C22H42O4

IUPAC Name:

bis(2-ethylhexyl) adipate

Details on test material:

- Name of test material (as cited in study report): di(2-ethylhexyl)adipate
- Physical state: clear, colorless liquid
- Analytical purity: 100.5 % for Lot. No. 0-62-494 and 101.4 % for No. GC-2-27-76 (determined by ester titration)
- Lot/batch No.: GC-2-27-76
- Storage condition of test material: at 4°C

Test animals

Species:

mouse

Strain:

B6C3F1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: NCI Frederick Cancer Research Center, Frederick, Maryland.
- Age at study initiation: 4 weeks old.
- Housing: 5 per cage in solid bottom suspended polycarbonate cages equipped with disposable nonwoven fiber filter sheets and Aspen-bed hardwood chips as bedding.
- Diet: powdered Wayne Lab Blox diet, ad libitum.
- Water: available via an Edstrom automatic watering system, ad libitum.
- Acclimation period: 2 weeks.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 - 31 °C.
- Humidity: 10 - 88 %.
- Air changes: 10 per hr.
- Photoperiod12 hrs dark /12 hrs light.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): at least every 14 days.
- Mixing appropriate amounts with (Type of food): chemical was mixed with aliquot of powdered Wayne Lab Blox animal feed, placing the mixture in a Petterson-Kelly twin-shell intensifier bar V-blender with the remainder of the feed and mix for 10 minutes.
- Storage temperature of food: 4 °C for no longer than 14 days.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The amounts of the test substance in 12500 and 25000 ppm samples were determined by vapour phase chromatography. One-gram samples of each of the above mixtures were triturated twice with 50-mL portions of methanol. The supernatant solutions were combined and diluted to a volume of 100 ml and analyzed. The mean of the analytical concentration was usually within 10 % of the theoretical.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
1600, 3100, 6300, 12500, 25000 ppm
Basis: nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
200, 387, 787, 1562, and 3125 mg/kg bw
Basis: actual ingested (recalculated based on food consumption). Only the corresponding value to 1600 ppm were given in the study report. The other doses were calculated with the mean food consumption used to calculate this given value.

No. of animals per sex per dose:

10

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: doses used are based on an acute study and a 14-day study (groups of five mice per sex were treated with five dose levels of the test substance in feed -up to 25000 ppm- for 14 days, followed by 1 day of observation with control diet. 1 group per sex were maintained as untreated controls. All surviving animals were killed after 15 days).

Examinations

Observations and examinations performed and frequency:

CAGE SIDE & CLINICAL OBSERVATIONS: yes
- Time schedule: twice daily

BODY WEIGHT: yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- no specifics given on how feed consumption was observed

Sacrifice and pathology:

GROSS PATHOLOGY: yes

HISTOPATHOLOGY: yes

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

mortality observed, treatment-related

Description (incidence):

1 female died (12500 ppm)

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

body weight depression in males (from 3100 ppm) and females (from 6300 ppm)

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

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

not examined

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
One mouse died as a result of an accident.

BODY WEIGHT AND WEIGHT GAIN
Weight gain depression was 10 % or more for male mice fed 3100 ppm or more. Weight gain depression was 10 % or more for female mice fed 6300 or 25000 ppm.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
No compound-related reduction in feed consumption was observed.

HISTOPATHOLOGY: NON-NEOPLASTIC
No compound-related histopathological effects were observed.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: decreased body weight gain at 387 mg/kg bw/day

2

Dose descriptor:

NOAEL

Effect level:

1 600 ppm

Based on:

test mat.

Sex:

male

Basis for effect level:

other: decreased body weight gain at 3100 ppm

3

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: decreased body weight gain

4

Dose descriptor:

LOAEL

Effect level:

3 100 ppm

Based on:

test mat.

Sex:

male

Basis for effect level:

other: decreased body weight gain

5

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: decreased body weight gain at 787 mg/kg bw/day

6

Dose descriptor:

NOAEL

Effect level:

3 100 ppm

Based on:

test mat.

Sex:

female

Basis for effect level:

other: decreased body weight gain at 6300 ppm

7

Dose descriptor:

LOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: decreased body weight gain

8

Dose descriptor:

LOAEL

Effect level:

6 300 ppm

Based on:

test mat.

Sex:

female

Basis for effect level:

other: decreased body weight gain

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

NOAEL (males, mice) = 200 mg/kg bw/day
LOAEL (males, mice) = 387 mg/kg bw/day
NOAEL (females, mice) = 387 mg/kg bw/day
LOAEL (females, mice) = 787 mg/kg bw/day

Executive summary:

A subchronic oral toxicity study similar to OECD 408 was performed with the structurally analogue substance in B6C3F1 mice at dose levels of 1600, 3100, 6300, 12500 and 25000 ppm for a period of 90 days. Ten animals per sex and dose received the test substance daily via diet, whereas a similar constituted control group was administered the plain diet.

No signs of toxic effects and no mortality were observed in any of the animals during the study period. An adverse decrease in body weight gain compared to controls was noted starting at 3100 ppm in males and at 6300 or 25000 ppm in females, respectively. Average food consumption was not altered between treated and control groups of both genders. No adverse effects were noted at histopathological examination while clinical chemistry and haematological parameters were not reported in this study.

Based on these results, a NOAEL of 1600 ppm was derived for male B6C3F1 mice, corresponding to an actual ingested dose of 200 mg/kg bw/day. NOAEL for female mice was found to be 387 mg/kg bw/day.