Distillation residue, butyl alcohols production, rectification

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

key study

Study period:

1995-05-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Repeated-dose toxicity studies performed with the only identified and quantified constituent of the registered substance, 2-ethylhexanol. Studies performed with Fischer F344 rats and B6C3F1 mice followed the current U.S. EPA Good Laboratory Practice Guidelines. In the absence of data on the registered substance these studies serve as a good surrogate studies: a) it represents the effects of the only identified and quantified constituent of the substance, since the constituents of this registered UVCB-substance cannot be recognized and its composition varies to the degree that composition cannot be fixed. b) studies have been performed following U.S. EPA Good Laboratory Practice. c) they provide information on effect levels of 2-ethylhexanol when administered via oral cavage.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1996

Report date:

1995

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

Principles of method if other than guideline:

Similar to that of OECD 408, but with 11 day sub-acute study was performed to adjust the proper doses for the actual 90-day study. Number of animals per treatment group per sex was 10.

GLP compliance:

yes

Remarks:

U.S EPA Good laboratory practice was adopted

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-ethylhexan-1-ol

EC Number:

203-234-3

EC Name:

2-ethylhexan-1-ol

Cas Number:

104-76-7

Molecular formula:

C8H18O

Details on test material:

- Name of test material (as cited in study report): 2-ethylhexanol
- Molecular formula (if other than submission substance): C8H18O
- Molecular weight (if other than submission substance): 130.23 g/mol
- Smiles notation (if other than submission substance): OCC(CC)CCCC
- Substance type: 2-Ethylhexanol (2-EH), or isooctanol, is a fatty alcohol, an organic compound
- Physical state: colorless liquid
- Analytical purity: 99,8%
- Purity test date: 99, 8%
- Stability under test conditions: concentration and homogeneity was checked by gas chromatography analysis of samples from each dose level at the start of 11-day studies and periodically in 13-week studies.
- Storage condition of test material: no data

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animal species: Fischer 344 rats and B6C3F1 mice

TEST ANIMALS
- Source:not disclosed, age: rats 33-35 days upon arrival, mice 43-44 days upon arrival.
- Age at study initiation: 42 days rats, 44 days mice
- Weight at study initiation: rats: males 103g (86-128) and females 81g (64-95), mice: males 23g (21-26) and females 19g (17-23)
- Fasting period before study: 16-20 hours
- Housing: following U.S. EPA GLP: rats housed singly in suspended stainless steel wire mesh cages and mice singly in plastic cages arranged in racks.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%):30-70
- Air changes (per hr): not disclosed
- Photoperiod (hrs dark / hrs light): 12 hours

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

polyoxyethylene glycerol triricinoleate, Cremophor EL, BASF Aktien Gesellshaft, Ludwigshafen, Germany

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): daily
- Mixing appropriate amounts with (Type of food): cavage solution in Cremophor LE
- Justification for vehicle choice: most stable dose formulation and minimized tract irritation and inflammation.

VEHICLE
- Justification for use and choice of vehicle (if other than water): not disclosed
- Concentration in vehicle:
Subacute: 11-day study 1000 and 1500 mg/kg bw ; 13-week study 25, 125, 250 and 500 mg/kg bw.
Oncogenicity: rats 50, 150, 500 mg/kg bw and mice 0, 50, 200 and 750 mg/kg bw.
- Amount of vehicle (if gavage): 10ml/kg bw or 1ml in mice 3ml in rats (solution of 0,005% Cremophor EL)
- Lot/batch no. (if required):not disclosed
- Purity: not disclosed

HOMOGENEITY AND STABILITY OF TEST MATERIAL: see test material details.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Gas chromatographic analysis of homogeneity and concentrations, start of the study (11-days study) and peridiocally (90-days study).

Duration of treatment / exposure:

11 days sub-acute study
90-days sub-chronic study

Frequency of treatment:

5-times a week, consecutively.

No. of animals per sex per dose:

11-day study: 10 females and 10 males, rats and mice
90-day study: 10 females and 10 males, rats and mice

Positive control:

not needed

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily treatment days, once non-treatment days.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: 11-day study: days -3, 0, 4, 10. 90-day study: day -1 and weekly thereafter

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food and water consumption were measured on 11-day study on days 4 and 10. In 90-day study average food consumption was determined weekly.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: 11-day study: retro-orbital bleeding prior to termination or after decapitation. In 90-days study: from fasted animals on the morning of days 29 and 84.
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- How many animals: no data
- Parameters checked in table [No.?] were examined: leucocytes, eryhtrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets and differential leucocytes, and reticulocytes (only 90-day study).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: 11-day study: retro-orbital bleeding prior to termination or after decapitation. In 90-days study: from fasted animals on the morning of days 29 and 84.
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- How many animals: no data
- Parameters checked in table [No.?] were examined.: alanine , aspartate aminotransferase, creatine kinase, gamma-glutamyltransferase, glucose, urea, total protein, albumin/globulin, Natrium, Potassium, Calcium, Chlorine, creatinine, cholesterol, triglycerides, total bilirubin.

URINALYSIS: Yes / No / No data
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: Yes / No / No data
- Animals fasted: Yes / No / No data
- Parameters checked in table [No.?] were examined.

Sacrifice and pathology:

Moribund animals were euthanized; dead and euthanized animals were immediately necropsied and assessed grossly. Gross and microscopic examinations were made as follows:
11-day study adrenals, brain, kidneys, livers, lungs, spleen, stomach, and testes were weighed and together with any gross lesions and thymuses were processed, fixed in 4% formalin, stained with hematoxylin.eosin and examined microscopically.
In 90-days study: drenals, brain, kidneys, livers, lungs, spleen, stomach, and ovaries from all animals were weighed with other tissues listed in U.S. EPA Health Effect guidelines (1978b) fixed in 4% formalin. Livers were also stained with oil red for lipid content and examined microscopically. Animals in 90-day ancillary studies were used only to determine hepatic peroxisome proliferation. Livers were removed at termination and weighed and cyano-insensitive pCoA activities and protein concentrations were determined.

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

Means and standard deviations were calculated for body weights, food and water consumption, clinical pathology results, and organ weights. Values for test groups were compared with controls in the main study by ANOVA followed by Dunnett's test and in ancillary studies by ANOVA followed by Student't t-test.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

reduced body weight

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

higher dose levels groups had reduced food intake 10-20%

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

increased liver, kidney, stomach and testes weights.

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Effect levels

Dose descriptor:

NOAEL

Effect level:

ca. 125 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects, clinical signs: reduced body weight and increased relative liver, kidney, stomach and testes weight.

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

These studies were peformed in order to derive suitable dose levels for oncogenicity testing (U.S EPA, 1987c) according to U.S EPA requirements. These studies were performed as an oral cavage since irritative effects of oesophagus and mucosa of the GI tract was wanted to be excluded as it is the primary effect on first contact with external surfaces of the body.

A no-effect level for target organ (liver) was established as 125 mg/kg day bodyweight for both rat and mouse.