3-methylbut-2-en-1-ol

Toxicity to reproduction

Administrative data

Endpoint:

toxicity to reproduction

Remarks:

other: sub-chronic

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP - Guideline study according to OECD 408 for repeated dose including reproductive parameters.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Guideline:

other: OECD 408 including reproductive parameters

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-methylbut-2-en-1-ol

EC Number:

209-141-4

EC Name:

3-methylbut-2-en-1-ol

Cas Number:

556-82-1

Molecular formula:

C5H10O

IUPAC Name:

3-methylbut-2-en-1-ol

Details on test material:

- Name of test material (as cited in study report): 3-Methylbut-2-en-1-ol (Prenol)
- Test substance No.: 00/0274-1
- Date of production: 10 Apr 2000
- Physical state / color: Liquid / colorless, clear
- Analytical purity: 99.1 % (method: gas chromatography)
- Homogeneity: Homogeneous
- Lot/batch No.: Ch. 00/18, Abl. Nr. 56-1706
- Stability under test conditions: Proven by reanalysis after the in life phase of the study.
- Storage condition of test material: Room temperature
- Other: The analyses were carried out at the Analytical Department of BASF Aktiengesellschaft.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Germany, (Strain: CRL:WI (GlX/BRL/HAN) IGS BR)
- Age at study initiation: 34 - 36 days
- Age at the start of the administration period: 41 - 43 days
- Range of weight at study initiation: males: 148.7 - 171.1 g (group mean: 159.2 g); females: 111.1 - 136.4 g (group mean: 123.5 g).
- Fasting period before study: No data
- Housing: singly
- Diet: Provimi KLIBA SA, Kaiseraugst / Switzerland, ad libitum
- Water: ad libitum
- Acclimation period: yes, 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24°C
- Humidity (%): 30 - 70%
- Air changes (per hr): fully air-conditioned room
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours

Administration / exposure

Route of administration:

oral: drinking water

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance was administered as a solution in drinking water. The appropriate amount of test substance was weighed, then drinking water was filled up to the desired volume and subsequently mixed using a magnetic stirrer. The test substance solutions were prepared twice a week.

VEHICLE
- Concentration in vehicle: 200, 1000 and 5000 ppm

STABILITY OF TEST SUBSTANCE IN VEHICLE: determined over a period of 4 days at room temperature at the start of the study. As the preparations were solutions in water, no homogeneity analyses were carried out. Concentration control analyses of the test substance preparations were performed in all concentrations at the start and the end of the administration period.

Details on mating procedure:

No mating performed

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the test substance in drinking water over a period of 4 up to 7 days at room temperature was determined before the start of the study.
Concentration control analyses of the test substance preparations were performed in all concentrations at the start and the end of the administration period. The recovery rates were within a range of 92.1 % - 101.1 % of the target concentrations.

Duration of treatment / exposure:

90 days

Frequency of treatment:

continuously

Details on study schedule:

No mating performed.

Doses / concentrations

Remarks:

Doses / Concentrations:
5000 ppm (calculated as 243.8 mg/kg bw/d in males, 307.2 mg/kg bw/d in females) 1000 ppm (calculated as 65.4 mg/kg bw/d in males, 82.1 mg/kg bw/d in females) 200 ppm (calculated as 14.4 mg/kg bw/d in males, 21.0 mg/kg bw/d in females).
Basis:
nominal in water

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

Post-exposure period: none
- Dose selection rationale: 5000 ppm as high concentration with expected toxic effects; 1000 ppm as mid concentration; 200 ppm as low concentration with no expected toxic effects.

Positive control:

No data

Examinations

Parental animals: Observations and examinations:

See chapter 7.5.1

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

At necropsy specimen were sampled from fasted anesthetized male animals in a randomized sequence for sperm analyses. Thereby, sperm analyses were performed towards the end of the administration period. Immediately after necropsy and organ weight determination the right testis and cauda epididymis were taken from all male animals.

The following parameters were determined. Thereby sperm motility examinations were carried out in a randomized sequence.
• sperm motility;
• sperm morphology;
• sperm head count (cauda epididymis);
• sperm head count (testis);

Litter observations:

Not examined

Postmortem examinations (parental animals):

GROSS NECROPSY
- The animals were sacrificed by decapitation under CO2 anesthesia. The exsanguinated animals were necropsied and assessed by gross pathology.
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS

ORGAN WEIGHT: Yes
Primary and secondary reproductive organ weights:
- testes,
- epididymides,
- ovaries,
- uterus,
- prostate gland.

For further information, see Chapter 7.5.1


HISTOPATHOLOGY: Yes
Primary and secondary reproductive organs:
Left testis, left epididymis and both ovaries (fixed in Bouin' solution; after fixation, the organs were embedded in paraplast).

The following organs were fixed in 4% formaldehyde solution:
- pituitary gland
- oviducts/uterus/vagina
- prostate gland, seminal vesicles
- female mammary gland

Organs were examinated in all animals of the control and the high dose group.

For further information, see Chapter 7.5.1

Postmortem examinations (offspring):

Not examined

Statistics:

For further information, see Chapter 7.5.1

Reproductive indices:

Not examined

Offspring viability indices:

Not examined

Results and discussion

Results: P0 (first parental generation)

Details on results (P0)

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
The estrous cycle was not examined in this study, therefore no data were available concerning this special reproductive function.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
No effects on reproductive organs (see organ weights below) and no treatment-related changes in sperm parameters were observed, up to the highest dose group (5000 ppm).
• Total spermatids/g testis
control: 123 ± 21.3;
200 ppm: 121 ± 15.0;
1000 ppm: 141 ± 15.1;
5000 ppm: 132 ± 19.4
• Total sperm/g cauda epididymis as listed below:
control: 519 ± 66.4;
200 ppm: 543 ± 92.8;
1000 ppm: 549 ± 84.1;
5000 ppm: 573 ± 105.5
• % normal sperm:
control: 98.4 ± 0.88;
200 ppm: 97.7 ± 1.01;
1000 ppm: 97.8 ± 0.89;
5000 ppm: 98.3 ± 1.32
• % abnormal sperm:
control: 1.6 ± 0.88;
200 ppm: 2.3 ± 1.01;
1000 ppm: 2.2 ± 0.89;
5000 ppm: 1.7 ± 1.32
• % mortility:
control: 89 ± 4.1;
200 ppm: 81 ± 6.3, p ≤ 0.01;
1000 ppm: 84 ± 6.9;
5000 ppm: 90 ± 4.8

The statistically significant changes in the dose group of 200 ppm were not dose dependent and are therefore assumed as not treatment related.
REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
The reproductive performance was not examined in this study.

ORGAN WEIGHTS (PARENTAL ANIMALS)
ORGAN WEIGHTS
5000 ppm (243.8 mg/kg bw/d in males, 307.2 mg/kg bw/d in females)
Concerning reproductive organs
• In female rats of the high dose group, the absolute mean weight of the ovaries was slightly although significantly decreased compared to controls (-13.6 %, p ≤ 0.05). The absolute mean ovary weights are tabulated in table 1 (see remarks on results).
• The relative ovary weights of the high dose females did not show statistically significant changes compared to the controls (see table 1).
• The mean relative weights of testes (+13 .7 %) and epididymides (+12 .7%) was increased compared to controls. This was regarded to be the secondary consequence of the decrease of the mean terminal body weight (-11 .7%).

Pathology (see below) did not indicate treatment-related gross lesions or microscopic findings in any of the organs investigated, thus providing no histopathological correlate to the observed changes in organ weights.

GROSS PATHOLOGY (PARENTAL ANIMALS)
- Concerning the reproductive organs of both sexes, there were no gross lesions noted.

HISTOPATHOLOGY (PARENTAL ANIMALS)
All microscopic findings recorded were either single observations, or they were recorded at a low incidence, or they occurred in control animals only, or at comparable incidence and graded severity in control and high dose animals.
Accordingly, no hisopathological correlate was obtained for the significantly increased mean relative weights of the testes and epididymides in male high dose group. No microscopic finding was obtained that may account for the significant decreased mean absolute weight of the ovaries.

For further information, see Chapter 7.5.1

Effect levels (P0)

1

Dose descriptor:

NOAEL

Effect level:

243.2 mg/kg bw/day

Sex:

male

Basis for effect level:

other: No effects on reproductive organs or sperm parameters observed in the highest dose which was examined. The NOAEL of 243.2 mg/kg bw corresponds to 5000 ppm.

2

Dose descriptor:

NOAEL

Effect level:

307.2 mg/kg bw/day (nominal)

Sex:

female

Basis for effect level:

other: No effects on reproductive organs observed in the highest dose which was examined. The NOAEL of 307.2 mg/kg bw corresponds to 5000 ppm.

Results: F1 generation

Details on results (F1)

No data

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Table 1: Absolute and Relative Mean Ovary Weights 

Dose Group	Mean Absolute Ovary Weights (mg ± SD)	Mean Relative Ovary Weights (mg ± SD)
Controls	94.9 ± 10.5	0.045 ± 0.005
200 ppm	91.5 ± 23.2	0.043 ± 0.012
1000 ppm	100.5 ± 16.1	0.050 ± 0.009
5000 ppm	82.0 ± 9.4  (p ≤ 0.05)	0.042 ± 0.004