Registration Dossier

Administrative data

Endpoint:
reproductive toxicity, other
Remarks:
Uterotrophic Bioassay
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from peer-reviewed journal

Data source

Reference
Reference Type:
publication
Title:
The estrogenic potential of salicylate esters and their possible risks in foods and cosmetics.
Author:
Zhaobin Zhanga, Chengxia Jia , Ying Hua, Libei Suna, Jian Jiao , Liang Zhaoa, Desheng Zhuc, Jun Li ,Yonglu Tianc, Huicheng Baic, Ruobao Li, Jianying Hu
Year:
2012
Bibliographic source:
Toxicology Letters 209 (2012) 146– 153

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD Test Guideline 440
Principles of method if other than guideline:
Uterotrophic Bioassay of 2-phenylethyl 2-hydroxybenzoate in mice
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- IUPAC name: 2-phenylethyl 2-hydroxybenzoate
- Common name: Phenethyl salicylate
- Molecular formula: C15H14O3
- Molecular weight : 242.2726 g/mol
- Smiles notation : c1ccc(cc1)CCOC(=O)c2ccccc2O
- InChl: 1S/C15H14O3/c16-14-9-5-4-8-13(14)15(17)18-11-10-12-6-2-1-3-7-12/h1-9,16H,10-11H2
- Substance type: Organic
- Physical state: Solid
Specific details on test material used for the study:
- IUPAC name: 2-phenylethyl 2-hydroxybenzoate
- Common name: Phenethyl salicylate
- Molecular formula: C15H14O3
- Molecular weight : 242.2726 g/mol
- Smiles notation : c1ccc(cc1)CCOC(=O)c2ccccc2O
- InChl: 1S/C15H14O3/c16-14-9-5-4-8-13(14)15(17)18-11-10-12-6-2-1-3-7-12/h1-9,16H,10-11H2
- Substance type: Organic
- Physical state: Solid

Test animals

Species:
mouse
Strain:
CD-1
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Experimental Animal Tech Co. of Weitonglihua (Beijing, China)
- Age at study initiation: Exact age was not mention immature mice were taken.
- Fasting period before study: No data available
- Use of restrainers for preventing ingestion (if dermal): No data available
- Housing: Animals were house in stainless steel wire-mesh cages
- Diet (e.g. ad libitum): basic diet, ad libitum
- Water (e.g. ad libitum): water ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12 h light: 12 h dark cycle

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

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
peanut oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: 3- fold serial dilution in peanut oil was used to prepare the doses from stock solution.

DIET PREPARATION
- Rate of preparation of diet (frequency): Daily
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): peanut oil
- Concentration in vehicle: 0, 11.1, 33.3, 100 and 300 mg/kg bw
- Amount of vehicle (if gavage): 5 mL/kg bw
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
not specified
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
3 days
Frequency of treatment:
daily
Doses / concentrations
Remarks:
0, 11.1, 33.3, 100 and 300 mg/kg bw
No. of animals per sex per dose:
Total: 60
0 mg/kg bw : 12
11.1 mg/kg bw: 12
33.3 mg/kg bw : 12
100 mg/kg bw : 12
300 mg/kg bw : 12
Control animals:
yes, concurrent vehicle
Details on study design:
not specified
Positive control:
17β-estradiol was used as the positive control( 10, 50 and 400 µg/kg bw/day)

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: No data available
- Time schedule: No data available
- Cage side observations checked in table [No.?] were included. Mortality observed

DETAILED CLINICAL OBSERVATIONS: No data available
- Time schedule: No data available

BODY WEIGHT: Yes
- Time schedule for examinations: Daily

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available
Estrous cyclicity (parental animals):
not specified
Sperm parameters (parental animals):
not specified
Litter observations:
not specified
Postmortem examinations (parental animals):
uteri weight were weighted.
Postmortem examinations (offspring):
not specified
Statistics:
Data are presented as the mean ± SD.
Reproductive indices:
not specified
Offspring viability indices:
not specified

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: estrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified

Details on results (P0)

Mortality: No effect on survival of treated female mice was observed as compared to control.

Body weight: No effect on body weight was observed in all the treated dose group as compared to control.

Organ weight: significant increase in uterine weight were observed at 33.3 mg/kg bw as compared to control. But when compared with 100 and 300 mg/kg bw not indicated statistical significance.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
mortality
body weight and weight gain
organ weights and organ / body weight ratios
other: No effect observed

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

Based on:
not specified
Sex:
not specified
Basis for effect level:
other: not specified
Remarks on result:
other: not specified

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F2 generation

General toxicity (F2)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not specified

Effect levels (F2)

Based on:
not specified
Sex:
not specified
Basis for effect level:
other: not specified
Remarks on result:
other: not specified

Target system / organ toxicity (F2)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

Number of mice used, number of mice that died during the experiment, arrival body weights and final body weights in the mouse uterotrophic assay. Data are presented as the mean ± SD.

Chemicals

Doses (mg/kg/day)

N (No. of deaths) 

Arrival weighta(g)

) Final weight (g)

E2

10µg/kg/day

12

7.71 ± 0.67

12.05 ± 1.37

 

50 µg/kg/day

12

7.49 ± 0.63

12.10 ± 0.96

 

400 µg/kg/day

11 (1)

7.65 ± 0.65

12.37 ± 1.14

Control

0

12

7.57 ± 0.65

12.02 ± 0.53

PES

11.1

12

7.66 ± 0.56

12.02 ± 0.42

 

33.3

12

7.69 ± 0.74

11.92 ± 0.80

 

100

12

7.67 ± 0.91

12.07 ± 0.72

 

300

12

7.67 ± 0.87

11.71 ± 0.57

aThe body weight on the day (PND 19) of arrival.

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 300 mg/kg bw for P generation when CD-1 female mice were treated with Phenethyl salicylate (IUPAC name: 2-phenylethyl 2-hydroxybenzoate) orally by gavage for 3 days.
Executive summary:

In a Uterotrophic Bioassay, CD-1 female mice were treated with Phenethyl salicylate (IUPAC name: 2-phenylethyl 2-hydroxybenzoate) in the concentration of 0, 11.1, 33.3, 100, 300 mg/kg/day in Peanut oil from PND 21 to PND 24. No effect on survival and body weight of treated female mice were observed as compared to control. Significant increase in uterine weight were observed at 33.3 mg/kg bw as compared to control. But when compared with 100 and 300 mg/kg bw not indicated statistical significance. Therefore, NOAEL was considered to be 300 mg/kg bw for P generation when CD-1 female mice were treated with Phenethyl salicylate (IUPAC name: 2-phenylethyl 2-hydroxybenzoate) orally by gavage for 3 days.