3-methyl-5-phenylpentanol

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

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

Adequacy of study:

key study

Study period:

1989-02-06 to 1989-03-20

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was performed in compliance with GLP, and to a method equivalent to the standardised guidelines OECD 474 and EU Method B.12 with some limitations were present in the study design. A different sampling regimen was used for the controls and lower dose to that recommended in the guideline, however the top dose used was compliant with the guideline, and no cytogenic effects were noted, the results were therefore considered suitable for assessment. The study was performed on a structural analogue of 3-methyl-5-phenylpentanol, the only difference between the two substances being the position of the methyl group. As such the results are considered to be representative of 3-methyl-5-phenylpentanol an accurate reflection of its cytogenicity.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

Sampling time for the lower dose groups and controls were only performed once. The highest dosing group third sample extended past 48 h post dosing.

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

25634-93-9

Cas Number:

25634-93-9

IUPAC Name:

25634-93-9

Constituent 2

Reference substance name:

2-methyl-5-phenylpentanol

EC Number:

405-890-8

EC Name:

2-methyl-5-phenylpentanol

IUPAC Name:

405-890-8

Test material form:

not specified

Details on test material:

- Storage condition of test material: Brown screw cap glass bottle

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SAVO-Ivanovas, 7964 Kisslegg
- Age at study initiation: 6-10 weeks
- Weight at study initiation: 25-30 g
- Assigned to test groups randomly: yes
- Housing: five per cage in Makrolon cages
- Diet: standard laboratory diet ad libitum
- Water: tap water ad libitum
- Acclimation period: at least five days prior to use

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): ca. 55 %
- Photoperiod (hrs dark / hrs light): Artificial light was provided in a 12/12 h light/dark cycle.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: arachis oil
- Amount of vehicle: 10 mL/kg

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The compound was dissolved in arachis oil. The solution was freshly prepared just prior to use.

Duration of treatment / exposure:

A single exposure

Frequency of treatment:

A single exposure

Post exposure period:

24, 48 or 72 h

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 250, 500 and 1000 mg/kg dosed as 10 mL/kg
Basis:
nominal conc.

No. of animals per sex per dose:

Five males and five females per dose

Control animals:

yes, concurrent vehicle

Positive control(s):

9,10-dimethyl-1,2-benzanthracene (DMBA) and cyclophosphamide

- Route of administration: DMBA was administered per os in olive oil and cyclophosphamide was dosed via intraperitoneal injection in phosphate buffered saline.
- Doses / concentrations: 50 mg/kg

Examinations

Tissues and cell types examined:

Bone marrow cells

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: An initial toxicity test demonstrated that the maximum tolerate dose was 1000 mg/kg

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Please refer to table 1 in the field "Any other information on materials and methods incl. tables".

DETAILS OF SLIDE PREPARATION:
The animals were killed by cervical dislocation. Bone marrow was removed from both femurs by rinsing with foetal calf serum. Bone marrow cells were centrifuged at 150 g for 10 minutes and the supernatant discarded. Smears were made from the pellet on slides and air dried according to haematological routine. Two slides were made per animal.

The preparations were stained using the May-Gruenwald-Giemsa method.
- Stained for 3 minutes in undiluted May-Gruenwald solution
- Stain for 2 minutes in May-Gruenwald, diluted with distilled water 1:1
- Rinse briefly in distilled water
- Stain for 10 minutes in Giemsa, diluted with distilled water 1:6
- Rinse thoroughly in distilled water
- Dry in air, clean the back of the slides with methanol
- Clear in xylene for 5 minutes, and mount in Eukitt

METHOD OF ANALYSIS: The slides were observed under a microscope with a 100x oil immersion objective lens at 1250 fold magnification. At least 1000 polychromatic erythrocytes per animal were scored for the incidence of micronuclei. The number of micronucleated normochromatic erythrocytes were also recorded. The ratio of polychromatic to normochromatic erythrocytes was determined for each animal by counting a total of 1000 erythrocytes.

Evaluation criteria:

If the ratio of polychromatic to normochromatic erythrocytes is determined and found to be significantly different from the control value, this is taken as an indication of cytotoxicity.

Statistics:

Statistical significance was determined according to the methods of Kastenbaum and Bowman (standard tables) (1970)

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

After dosing, the animals of all groups treated with the test substance showed reduced motility. The most pronounced effect in the highest dose group.

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RESULTS OF STUDY
- Induction of micronuclei (for Micronucleus assay): None of the different treatment times and dose levels studied produced any increase in the frequency of micronucleated polychromatic erythrocytes.
- Ratio of PCE/NCE (for Micronucleus assay): The ratio of polychromatic to normochromatic erythrocytes was normal
- Appropriateness of dose levels and route: The maximum tolerated dose was used as determined by the range finder test (details of which were not included as part of the report)

Any other information on results incl. tables

Table 2: Results

Dose (mg/kg)	Sampling time	Sex	No. of animals	Ratio PE/NE	Micronucleated PE (0/00)
					Mean ± S.D.	(Min/Max)
Control	24 h	Male	5	1.45	1.59 ± 1.02	(0.00/3.00)
		Female	5	1.57	1.79 ± 0.75	(0.99/2.99)
250	24 h	Male	5	1.11	1.96 ± 0.62	(0.99/2.95)
		Female	5	1.18	0.98 ± 0.62	(0.00/1.96)
500	24 h	Male	5	1.39	1.37 ± 0.50	(0.89/1.98)
		Female	5	1.07	1.57 ± 0.48	(0.99/1.99)
1000	24 h	Male	5	1.61	2.38 ± 1.01	(0.98/3.96)
		Female	5	2.02	1.39 ± 1.01	(0.00/2.98)
1000	48 h	Male	5	1.45	1.39 ± 1.01	(0.00/2.98)
		Female	5	1.57	1.39 ± 0.80	(0.00/1.99)
1000	48 h	Male	5	1.81	1.97 ± 1.09	(0.99/3.98)
		Female	5	1.98	1.39 ± 1.02	(0.00/2.99)
DMBA 50	48 h	Male	5	1.20	12.16 ± 1.33	(9.98/14.00)
		Female	5	1.06	9.55 ± 2.32	(6.94/12.90)
CP 50	24 h	Male	5	1.08	24.65 ± 3.87	(18.88/28.36)
		Female	5	0.99	29.98 ± 3.07	(25.74/33.46)
DMBA: 9,10-dimethyl-1,2-benzanthracene CP: cyclophosphamide PE: polychromatic erythrocytes NE: normochromatic erythrocytes S.D.: Standard deviation p.o.: Administration per os i.p.: intraperitoneal injection

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Under the conditions of the test, it was concluded that the test substance did not induce chromosomal damage or damage to the mitotic apparatus in bone marrow cells of mice. The study was performed on a structural analogue of 3-methyl-5-phenylpentanol, the only difference between the two substances being the position of the methyl group. As such the results are considered to be representative of 3-methyl-5-phenylpentanol and an accurate reflection of its cytogenicity.

Executive summary:

In a GLP compliant study, conducted using a method similar to standardised guidelines OECD 474 and EU Method B.12 (with some minor deviations), the cytogenicity of the test substance was determined in male and female NMRI mice. The study was performed on a structural analogue of 3-methyl-5-phenylpentanol, the only difference between the two substances being the position of the methyl group. As such the results are considered to be representative of 3-methyl-5-phenylpentanol and an accurate reflection of its cytogenicity. Under the conditions of the test, it was concluded that the test substance did not induce chromosomal damage or damage to the mitotic apparatus.