Registration Dossier

Administrative data

Endpoint:
genetic toxicity in vivo
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Data is from study report (NTRL)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993

Materials and methods

Principles of method if other than guideline:
Dominant Lethal Assay was performed to evaluate the mutagenic nature (in vivo) of the test compound 3-Phenoxybenzyl alcohol on SD rats
GLP compliance:
not specified
Type of assay:
rodent dominant lethal assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
- Name of test material (as cited in study report): 3-Phenoxybenzyl alcohol (T1375)
- Molecular formula (if other than submission substance): C13H12O2
- Molecular weight (if other than submission substance): 200.236 g/mol
- Substance type: Organic
- Physical state: Liquid
Purity No data available
- Impurities (identity and concentrations): No data available

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
Male: Charles River, .Wilmington, Massachusetts
Females: Flow Laborato-ries, Dublip, Virginia
- Age at study initiation: 8-10 week old
- Weight at study initiation:
Males: 225-260 g
Females: 185- 215 g
- Assigned to test groups randomly: No data available
- Fasting period before study: No data available
- Housing: Housed in cages
- Diet (e.g. ad libitum): standard laboratory diet ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 10-14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%): No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

IN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: Corn oil
- Justification for choice of solvent/vehicle: No data available
- Concentration of test material in vehicle: 238.0, 23.8 or 2.38 mg/Kg
- Amount of vehicle (if gavage or dermal): No data available
- Type and concentration of dispersant aid (if powder): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on exposure:
For oral route
PREPARATION OF DOSING SOLUTIONS: No data available

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available Prepared fresh prior to use and used for five days
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available
Duration of treatment / exposure:
Duration of exposure: 5 days
Frequency of treatment:
Daily
Post exposure period:
7 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
238.0, 23.8 or 2.38 mg/Kg
Basis:

No. of animals per sex per dose:
Male: 10/group : 30
Females: 2/mating with 1 male/week : 420
Control animals:
yes, concurrent vehicle
Positive control(s):
Positive controls
Triethylene melamine (TEM)
- Justification for choice of positive control(s): No data available
- Route of administration: No data available
- Doses / concentrations: 0.5 mg/Kg bw

Examinations

Tissues and cell types examined:
Corpora lutea, uterine horns, fetal death, total no. of implantations from females
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: LD5, LD0.5 and LD0.05

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Fourteen days from the mid-week of mating

DETAILS OF SLIDE PREPARATION:

METHOD OF ANALYSIS: Fourteen days from the mid-week of mating, the females were sacrificed by C02 asphyxiation and the abdominal cavity was exposed. The membrane was removed from each ovary and the corpora lutea for each ovary were counted and recorded separately. In addition, both uterine horns were examined and fetal deaths and total implantations were determined and recorded separately for each horn.

OTHER: No data available
Evaluation criteria:
Females mated with negative control males must show a total of 8-15 implantations.

Females mated to positive control males must exhibit severe fetal damage. There must be a statistically significant reduction in implantations and there must be a statistically significant increase in females with 2 or more dead implants. This damage must be seen between weeks 2 and 7 of the spermatogenic cycle. A test which does not meet these requirements must be repeated.
Statistics:
1. The t-test was used to compare pairwise the means of each treated group with the means of each control (negative control, historic control, positive control), and the types of controls between themselves within each week. Each comparison was considered to be between two independent, random samples of unequal variance, and a significant difference in either direction (2-sided) between the means of each sample was being sought.

2. Chi-square analysis was used to ascertain significant relationships between the data classified by one factor into one of two rows and by a second factor into one of two columns. In addition, the negative and positive controls were compared to each other. All comparisons were done within each week.

3. Analysis of regression was used to detect a dose response relationship between the parameter and the arithmetic or logarithmic dose.

4. The analysis of linear trend in proportion was used to detect a dose response relationship between the parameter and the arithmetic or logarithmic dose.

5. Analysis of variance is a technique of partitioning the total variation in a set of data into a number of components (sums of squares) which can be assigned to main effects (e.g., groups, weeks), interactive
effects (e.g.; groups by weeks, males within groups by weeks), or error.

6. The probit analysis is a technique used in biological assays based on a quantal response, i.e. a response which has an all-or-none character. Using several statistical transformation techniques, a sufficiently
accurate weighted probit regression line was obtained and was used to calculate the effective doses and their fiducial limits at various levels of response.

7. The Freeman-Tukey arcsine transformation for a proportion is defined by the following formula, where X is the number of successes, N is the number of trials, and θ (theta) is the transformed value.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: The LD50 (1668 mg/kg body weight) provided by Ethyl Corporation was used to calculate the LD5, LD0.5 and LD0.05
- Solubility: No data available
- Clinical signs of toxicity in test animals: Mortality
- Evidence of cytotoxicity in tissue analyzed: No data available
- Rationale for exposure: No data available
- Harvest times: No data available
- High dose with and without activation: No data available
- Other: No data available

RESULTS OF DEFINITIVE STUDY
- Types of structural aberrations for significant dose levels (for Cytogenetic or SCE assay): No data available
- Induction of micronuclei (for Micronucleus assay): No data available
- Ratio of PCE/NCE (for Micronucleus assay): No data available
- Appropriateness of dose levels and route: No data available
- Statistical evaluation: No data available

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
3-Phenoxybenzyl alcohol does not induce dominant lethal mutation in mice and is negative for mutation in vivo.
Executive summary:

Dominant Lethal Assay was performed to evaluate the mutagenic nature (in vivo) of the test compound 3-Phenoxybenzyl alcohol on Sprague Dawley rats.

 

Three groups of male rats, 10 per group, were given five intragastric treatments with the test agent at levels of LD5, LD0.5and LD0.05from Monday through Friday. The following week each male was then mated with 2 females from Monday through Friday. On Friday, the females were removed from the cage and the male was allowed to rest for 2 days. On the subsequent Monday, the male was mated with 2 new females and the process repeated until each male had been mated for 7 weeks with 2 new females per week.

 

Fourteen days from the mid-week of mating, the females were sacrificed by CO2asphyxiation and the abdominal cavity was exposed. The membrane was removed from each ovary and the corpora lutea for each ovary were counted and recorded separately. In addition, both uterine horns were examined and fetal deaths and total implantations were determined and recorded separately for each horn.

 

The test compound 3-PBA appeared to exhibit minimal activity in number of total implantations, preimplantation losses, and number of live implants per pregnant female. However, the effect on total or live implants per pregnant female occurred at a single dose level at week 3 and 7. The variation in corpora lutea counts does not reflect any biological activity of the doses tested but rather is a variation in the female animals. The increase seen in pre-implantation losses occured at a single dose at week 1 only. No statistically significant dose response was observed.

 

The data suggest that the test compound exhibits little or no mutagenic activity in the dominant lethal test.