Acetone

Toxicity to reproduction

Administrative data

Endpoint:

reproductive toxicity, other

Remarks:

other: toxicity study with specific investigation of effects on male fertility

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study meets basic scientific principles as a study of effects on male fertility with the following restictions: as the acetone-treated groups were part of a study on the combined action of acetone and DEHP at fixed acetone concentrations only a single acetone concentration was used for each exposure period, negative results not presented in detail; study acceptable for weight of evidence approach

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

publication

Title:

Toxicity study of di(2-ethylhexyl)phthalate (DEHP) in combination with acetone in rats

Author:

Dalgaard M, Ostergaard G, Lam HL, Hansen EV, Ladefoged O

Year:

2000

Bibliographic source:

Pharmacol Toxicol 86: 92-100

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Assessment of male fertility via reproductive indices (matings after 3 w pretreatment), immunohistochemistry of testes, and histopathology of male reproductive organs; systemic toxicity assessed by body weights, clinical biochemistry parameters, behavioural toxicity, organ weights and histopathology

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Acetone

EC Number:

200-662-2

EC Name:

Acetone

Cas Number:

67-64-1

Molecular formula:

C3H6O

IUPAC Name:

propan-2-one

Details on test material:

- Analytical purity: analytical grade (no further data)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Möllegaard Breeding Centre Ltd., DK-4623 Ll. Skensved, Denmark
- Age at study initiation: Females 10 weeks
- Weight at study initiation: Males: 160 g
- Housing: 2 rats/cage
- Diet: ad libitum
- Water: libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +- 1
- Humidity (%): 55 +- 5
- Photoperiod (hrs dark / hrs light): 12/12 (reversed day / night cycle)

Administration / exposure

Route of administration:

oral: drinking water

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 1 week during the 4th week of exposure
- Proof of pregnancy: vaginal plug
- After successful mating each pregnant female was caged for 15 days

Duration of treatment / exposure:

4 weeks: 1.0 %-dose group
9 weeks: 0.5 %-dose group

Frequency of treatment:

daily

Details on study schedule:

During the last week of the 4-week study all male rats were mated with undosed females. A female rat was placed in a cage with a male rat for 6 h daily until mating was recognised (presence of vaginal plug) or for the maximum of 7 days.
On day 15 after mating female rats were sacrificed and the uteri were examined for the presence of implantations and foetuses.

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0, 5000 mg/L (0.5 %)
Basis:
nominal in water
9 week exposure, corresponding to an estimated dose of 650 mg/kg bw/d

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0, 10000 mg/L (0, 1 %)
Basis:
nominal in water
4 week exposure; corresponding to a dose of 1300 mg/kg bw/d

No. of animals per sex per dose:

10 treated males per dose and exposure duration
10 untreated females per male dose group (only in the 4 week study)

Details on study design:

- Further dose groups exposed to DEHP as the intention of the study was to investigate the combined toxicity of DEHP and acetone.

Positive control:

- Dose groups treated with DEHP can be regarded as positive control groups.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Time schedule for examinations: weekly for each cage (2 animals)

WATER CONSUMPTION : Yes
- Time schedule for examinations: weekly for each cage (2 animals)

OTHER: Yes
CLINICAL BIOCHEMISTRY: blood samples taken on day before sacrifice from orbital plexus. Plasma was isolated and analysis was performed on a COBAS-MIRA autoanalyzer by use of commercially available test kits. Clinical biochemical parameters examined were: alanine aminotransferase ALAT, bilirubin, cholesterol, glucose, triglycerides, creatinine, urea, protein, uric acid, lactate dehydrogenase LDH

BEHAVIOUR: Rats observed in Functional Observation Battery during handling and in an open field for a number of sensory, motor and physiological endpoints, for responsiveness to visual, auditory and tactile stimuli. Additionally, foot splay, righting reflex and fore- and hindlimb grip strength were measured.

Sperm parameters (parental animals):

Histopathology of testes, epididymis, seminal vescicles
grading of histopathological findings of the testes: normal, slight/moderate atrophy, severe atrophy

Expression of the cell structure protein vimentin: immunohistochemisty in Sertoli cells, surrounding basal lamina pria and in interstitial cells

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals after termination of exposure
- Maternal animals: on day 15 after mating

ORGAN WEIGHTS (male rats): liver, kidneys, adrenals, heart, spleen, brain, testes, epididymis, seminal vescicles

HISTOPATHOLOGY
- Male rats: light microscopic examination of liver, kidneys, adrenals, heart, spleen, brain, male reproductive organs (see Sperm parameters)

Statistics:

two-factor ANOVA: FOB test, food and water consumption, clinical biochemistry, body and organ weights
Chi-square test: fertility data

Reproductive indices:

- Number of males without recognised mating
- Number of pregnant females
- Number of implantations
- Number of dead or retarded foetuses

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function / performance (P0)

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL BIOCHEMISTRY
- Glucose level in plasma (mmol/l): significant decrease in 1 % acetone-treated rats (p<0.05); control 10.4 mmol/L, 1 % acetone 8.34 mmol/L

BEHAVIOUR
- Hindlimb grip strength: significant decrease in 1 % acetone-treated rats (p<0.05); control 820, 1 % acetone 650
- Forelimb grip strength: decrease in 1 % acetone-treated rats not significant; control 1080, 1 % acetone 890

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
- Number of males without recognised matings: control 1/10, 1 % acetone 0/10
- Number of pregnant females: control 9/10, 1 % acetone 9/10

Effect levels (P0)

1

Dose descriptor:

NOEL

Effect level:

10 000 mg/L drinking water

Sex:

male

Basis for effect level:

other: male fertility

2

Dose descriptor:

LOAEL

Effect level:

10 000 mg/L drinking water

Sex:

male

Basis for effect level:

other: other: behavioural effect

Results: F1 generation

Effect levels (F1)

Remarks on result:

other: Not investigated, specific study on male fertility

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Acetone exposure (5000 mg/L drinking water for 8 weeks or 10000 mg/L drinking water for 4 weeks) had no adverse effect on male fertility.

Executive summary:

Male Wistar rats were exposed to 0 or 10,000 mg acetone/L drinking water for 4 weeks or 0 or 5,000 mg acetone/L drinking water for 9 weeks (corresponding to doses of 1,300 or 650 mg/kg bw/d). Clinical signs, body weight, food and water consumption, organ weights, and clinical biochemical parameters were examined. Behavioural toxicity was screened via a Functional Observation Battery. In the last week of the 4 -week study each male was mated with an untreated female rat to investigate reproductive performance (number of males without recognised mating, number of pregnant females, number of implantations, number of dead or retarded fetuses). Histopathology was performed on liver, kidneys, adrenals, heart, spleen, brain, and with special emphasis on male reproductive organs (testes, epididymis, seminal vescicles). Expression of the cell structure protein vimentin was analyzed via immunohistochemistry in the testes in Sertoli cells, the surrounding basal lamina propria and in interstitial cells.

Acetone exposure had no adverse effect on male fertility (NOEL 10,000 mg/L). Four weeks of exposure to the 10,000 -mg/L dose level exerted effects outside the reproductive system as the glucose level was decreased in plasma and parameters of behavioural toxicity were changed. Hindlimb and forelimb grip strengths were decreased with a significant effect on the former parameter only.