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Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
Not reported
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was performed in line with GLP and the method was designed in line with good scientific principles. The study predates the inception of the OECD guideline 416 (2-generation reproductive toxicity), but is in basic compliance with the current standardised guideline. There is a good level of detail in reporting, sufficient to assess the quality of the data. The read-across approach has been used since phenoxyethanol is structurally similar to the test material but is more toxicologically active. The results presented are therefore taken to be the worst case scenario.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Principles of method if other than guideline:
The reproductive toxicity of the test material was investigated in Swiss CD-1 mice. Mice were exposed to the test material in diet at concentrations of 0, 0.25, 1.25 and 2.5% in diet equivalent to 0, 0.4, 2.0 and 4 g/kg bw/day (a range finding study was conducted with concentrations of 0, 1, 2.5, 5, 7.5 and 10% in feed for 2 weeks). Observations during the study included general toxicity, sperm analysis, reproductive performance, pup abnormalities and assessment of possible sex related effects (crossover mating).
GLP compliance:
yes
Limit test:
no
Species:
mouse
Strain:
CD-1
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: (P) 11 wks at the beginning of the continuous breeding test; (F1) 74 ± 10 days.
- Housing: Animals were housed by sex in solid bottom polypropylene or polycarbonate cages with stainless steel lids during the pre-mating period. The animals were then housed either individually or in breeding pairs.
- Diet: ad libitum.
- Water: Deionized water available ad libitum.
- Acclimation period: 2 weeks.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2 °C.
- Photoperiod (hrs dark / hrs light): 14 hours light (10 hours dark).
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet: Weekly (diet was shown to lose approximately 5 % over 7 days).
- Mixing appropriate amounts with diet: Each dose level was blended into a small amount of ground diet, which was then added to a pre-weighed portion and mixed.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 98 days as breeding pairs in the continuous breeding phase, in the crossover trial and the offspring fertility assessment animals were cohabited for 7 days
- Proof of pregnancy: vaginal plug
- After successful mating each pregnant female was caged (how): In the offspring fertility test, animals were housed individually after the cohabitation period. During the continuous breeding phase, the animals cohabited for 98 days.
- Any other deviations from standard protocol: In the crossover mating trial, the parental animals from the continuous breeding phase were cross mated with control animals to determine the affected sex. Three groups of 20 pairs were used: control males x control female; high dose males x control females; control males x high dose females. In the offspring fertility check, the last litter produced from the continuous breeding phase and cohabited with a mate from a different litter within the same dose group.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
No details of the methods used were reported. Aliquots of six representative samples were analysed during the study, the concentration was shown to be within 96 - 105% of the nominal concentration.
Duration of treatment / exposure:
- Range finding test: 14 days.
- Continuous breeding study: 7 day pre-mating dosing followed by 98 days cohabitation.
- In the cross-over mating trial parental animals selected from the continuous breeding study were necropsied after 3 weeks.
- The animals selected for the offspring fertility test were necropsied at the end of the test (after delivery).
Frequency of treatment:
Daily, dosed feed was available ad libitum, apart from the 7 days were breeding pairs were cohabited in the cross-over mating trial.
Details on study schedule:
- F1 parental animals not mated until 74 ± 10 days after selected from the F1 litters.
- Selection of parents from F1 generation when pups were 21 days of age.
Remarks:
Doses / Concentrations:
4000, 2000, 400 and 0 mg/kg bw /day
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
0, 0.25, 1.25 and 2.5%
Basis:
actual ingested
No. of animals per sex per dose:
- 8 animals/sex/group in each of the range finding dose levels
- 40 breeding pairs in the control group and 20 breeding pairs per dose were using in the continuous breeding phase of the study.
- In the crossover mating test 20 breeding pairs per group (3 in total) were used.
- In the offspring fertility test, 8 to 10 litters from the F1 generation from the control group and the 1.25 % dose level were selected for rearing to sexual maturity. At 74 ± 10 days one to three female and male pups were selected from each surviving litter and selected randomly for breeding. 19 breeding pairs per dose were used in this test.
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: A 14-day range finding study was included in order to select an appropriate dose for the study. Five dose groups and a control were exposed to 10, 7.5, 5.0, 2.5, 1.0 and 0.0 % in diet for two weeks. Animals were assessed for clinical signs, bodyweight changes and food consumption. Based on the results of this study, the appropriate doses were selected for the continuous breeding study. As both male and female animals lost 10 % of their bodyweight in the 7.5 and 10 % groups; 0, 0.25, 1.25 and 2.5 % were selected as the dosing levels in the continuous breeding trial.
Parental animals: Observations and examinations:
All animals (continuous breeding study, crossover mating trial and the offspring fertility test) were assessed for the following.

CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes calculated as the average g/day/mouse
Sperm parameters (parental animals):
The following parameters examined in all male mice in all tests within the study; sperm count, sperm motility and sperm morphology.

In addition, the following were examined in the continous breeding test and the offspring fertility test (F0 and F1) animals; testis weight and epididymis weight.
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 and F2 offspring; number and sex of pups, stillbirths, live births, postnatal mortality, weight gain and fertility.

The number of litters produced by the F0 generation in the continuous breeding phase of the study was also evaluated.

GROSS EXAMINATION OF DEAD PUPS: no
Postmortem examinations (parental animals):
ORGAN WEIGHTS
In the male animals from the continuous breeding study, the females from the crossover mating test and both the males and females from the offspring fertility test the liver weights were recorded.

In addition the following organs and tissues from male animals in the continuous breeding test and the offspring fertility test were weighed: right epididymis, right testis, seminal vesicles and prostate.
Postmortem examinations (offspring):
ORGAN WEIGTHS
As listed above, the following organ weights from the pups produced in the continuous breeding study used in the offspring fertility test were examined: liver, right epididymis, right testis, seminal vesicles and prostate.
Statistics:
The findings from the continuous breeding study were assessed using the Cochran-Armitage test to determine whether there was a dose-related trend. In the cross-over mating trial, as it was not possible to test for a dose-related trend, the findings were assessed using a Chi squared test for homogeneity to determine the overall difference in fertility among groups. Fisher’s exact test was As a pairwise comparisons between the control and dosed groups.
Dose groups means for sex ratio and proportion of live pups were assessed using the Kruskal-Wallis test to examine overall difference and Jonckheere’s test to assess ordered differences. Pairwise comparisons of treatment group means were performed using the Wilcoxon-Mann-Whitney U test.

To prevent the number of pups per litter affecting the assessment of average pup weight an analysis of covariance was used, using the average litter size and the covariate including live and dead pups. Least-squares estimated of dose group means, adjusted for litter size were tested using an F test to assess overall equality and a t test to test for pairwise equality. To prevent interference from potential sex differences, these were performed with male and females separately and also both sexes combined.

An analysis of covariance was also used to adjust organ weights appropriately for bodyweight. Unadjusted body and organ weights were analysed by the Kruskal-Wallis and the Wilcoxon-Mann-Whitney U tests. Jonckheere’s test was used to test for dose-related trends.
Reproductive indices:
The following reproductive indices were calculated using equations below:

Mating index (%) = (Number of animals with copulatory plug/Number of animals cohabited breeding pairs) x 100

Fertility index (%) = (Number of fertile pairs/Number of animals with copulatory plug) x 100
Offspring viability indices:
The number of litters and live pups per litter were assessed per fertile breeding pair from this figure the treatment group means were determined. The proportion of live pups was defined as the number of pups born alive divided by the total number of pups produced by each breeding pair. The sex ratio was expressed as the proportion of male pups born alive out of the total number of live pups born to each fertile pair.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Three females died in the continuous breeding study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Males only.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Males only.
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
not examined
Other effects:
no effects observed
Description (incidence and severity):
Test substance intake: No treatment related effects.
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
No treatment related effects.
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
2.5 % EGPE in diet reduced the number of litters delivered per breeding pair and reduced litter size and number of live births.
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
The test substance was found to have a low general toxicity, with only three female dying in the continuous breeding test.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
In the continuous breeding study, only a minimal effect was noted on male bodyweight (2 % decrease) with no change on average in females. There was no significant effect on feed consumption. The approximate daily consumption was 5.6 g/day/mouse. At necropsy at the end of the cross-over mating test, there was a significant decrease in the bodyweight for males exposed to 2.5 % EGPE.

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
There were no effects on feed consumption. The average daily test substance intake was approximately 400, 200 and 4000 mg/kg bw/day from the 0.25, 1.25 and 2.5 % groups respectively. The test substance intake varied in the females with the stage of gestation due to fluctuations in bodyweight.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
In the Cross-over mating test there were no differences between the treated and control males in respect to sperm concentration, percentage of motile sperm and percentage of abnormal sperm.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Continuous exposure in the continuous breeding test did not affect the number of breeding pairs able to produce at least one litter. Exposure to 2.5 % of EGPE was found to reduce the number of litters delivered per breeding pair and significantly reduced the size and proportion of pups born alive. Further assessment of the litters demonstrated that in the high-dose group only 12 out of 20 pairs (60 %) had a fifth litter compared to 36 out of 40 (90 %) in the control. The number of pups per litter was not found to be affected. In the cross-over mating test, the results did not show which sex was affected by administration of the test material. The only difference was live pup weight was significantly decreased by 12 % in the control males crossed with the 2.5 % dosed females.

ORGAN WEIGHTS (PARENTAL ANIMALS)
In the cross-over mating trial, when adjusted for bodyweight, the liver weight was found to be significantly increased in both males and females (21 and 60 % respectively). There were no significant difference in weight of the right testis, prostate and epididymis between controls and the 2.5 % EGPE males.
Dose descriptor:
NOAEL
Effect level:
400 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: body weight, liver weight and number of litters produced, live pup weight and litter size
Clinical signs:
not examined
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
High lethality was seen in the 2.5 % group.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Dose related decrease in body weight
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No treatment related effects.
Gross pathological findings:
not examined
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
Exposure to 2.5 % of EGPE in the continuous breeding test was found to reduce the number of litters delivered per breeding pair and significantly reduced the size and proportion of pups born alive. Further assessment of the litters demonstrated that in the high-dose group only 12 out of 20 pairs (60 %) had a fifth litter compared to 36 out of 40 (90 %) in the control. The number of pups per litter was not found to be affected. In the final litters produced by the continuous breeding test, pup lethality was pronounced during the lactation and post-weaning periods in the 1.25 and 2.5 %. By day 21 (weaning) only eight litters in the 1.25 and 2.5 % dosing groups had sufficient pups for the offspring fertility test. From birth to the start of mating, 12 of 87 (14 %) of the selected offspring for the fertility test in the control group, 20 of 113 pups (18 %) in the 0.25 % EGPE group, 33 out of the 84 pups (39 %) in the 1.25 % EGPE dose group and 66 out of the 76 pups (87 %) in the 2.5 % EGPE treatment group had died. Due to the high lethality in the 2.5 % group (25 of 32 males and 21 of 24 females), the offspring fertility test was performed with the pups from the 1.25 % dosing group.

BODY WEIGHT (OFFSPRING)
There was a significant dose related decrease in adjusted live pup weight during the continuous breeding test. In the cross-over mating test live pup weight was significantly decreased by 12 % in the control males crossed with the 2.5 % dosed females. The F1 pups selected for the offspring fertility test (the last litters produced by the continuous breeding test) were found to have dose-related decreased bodyweights at birth, weaning and at mating (the start of the offspring fertility test). At the end of the offspring fertility test, bodyweights were found to be decreased in both males and females dosed with 1.25 % EGPE.

SEXUAL MATURATION (OFFSPRING)
In the offspring fertility test, there were no statistically significant effects on the proportion of copulatory plug positive matings (mating index), fertile pairs (fertility index), pups born alive, or number of pups per litter when compared to the control pairs. Consistently, the live pup weights for the F2 generation were reduced. The offspring fertility test did not indicate any effects on sperm concentration, percentage of motile sperm or percentage of abnormal sperm.

ORGAN WEIGHTS (OFFSPRING)
The F1 pups used in the offspring fertility test were found to have increased liver weights when adjusted for bodyweight at the end of the test. There was no effect on right testis, prostate or epididymal weight. Seminal vesicle weight was found to be significantly decreased compared to controls.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
400 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: body weight, liver weight and live pup weight
Reproductive effects observed:
not specified

Table 1: Reproductive performance

Parameter

Continuous Breeding Test

Crossover Mating Test

Offspring Fertility Test

Control

0.25% EGPE in diet

1.25% EGPE in diet

2.5% EGPE in diet

Control male x Control female

2.5 % treated male x control female

Control male x 2.5 % treated female

Control

1.25% EGPE in diet

No. with copulatory plugs/No. cohabited

 

18/18

18/20

14/17

18/19

19/19

Mating index (%)

100

90

82

95

100

No. fertile/No. with copulatory plugs

38/38

20/20

19/20

18/18

16/18

16/18

12/14

17/18

18/19

Fertility index (%)

88

88

86

94

95

Litters per breeding paira

4.8 ± 0.1

4.8 ± 0.1

5.0 ± 0.1

4.5 ± 0.2

Live pups per littera

11.0 ± 0.4

11.6 ± 0.5

11.3 ± 0.5

9.0 ± 0.8*

9.06 ± 1.01 (16)

9.75 ± 0.90 (16)

7.42 ± 1.03 (12)

11.1 ± 0.5 (19)

10.7 ± 0.4 (19)

Proportion of pups born alivea

0.98 ± 0.01

0.99 ± 0.01

0.98 ± 0.02

0.90 ± 0.60*

0.86 ± 0.09 (16)

0.96 ± 0.02 (16)

0.92 ± 0.06 (12)

0.98 ± 0.01 (19)

0.99 ± 0.01 (19)

Live pup weight (g)a

1.65 ± 0.02

1.59 ± 0.03

1.58 ± 0.02

1.52 ± 0.04*

Adjusted live pup weight (g)ab

1.65 ± 0.02

1.61 ± 0.02

1.59 ± 0.02*

1.48 ± 0.03*

1.71 ± 0.04 (14)

1.72 ± 0.04 (16)

1.51 ± 0.04 (12)*

1.55 ± 0.02 (19)

1.45 ± 0.02 (19)*

* Significantly different from control group (P < 0.05)

aMean ± SE. Number of observations included in parentheses were appropriate

bMeans adjusted for total number of live and dead pups per litter by analysis of covariance

 

Table 2: Bodyweights, organ weights and sperm parameters of male mice in the continuous breeding study and male mice in the offspring fertility test

Observationa,b

Continuous Breeding Study

Offspring Fertility Test

Control

2.5 % EGPE in feed

Control

1.25% EGPE in feed

Number of males

37

20

20

20

Bodyweight (g)

38.81 ± 0.59

36.63 ± 0.58*

34.61 ± 0.73

30.71 ± 0.59*

Organ weights

Liver (g)

2.05 ± 0.03

2.48 ± 0.05*

1.85 ± 0.04

2.05 ± 0.04*

Right epididymis (mg)

64 ± 4

54 ± 6

50 ± 1

43 ± 1

Right testis (mg)

135 ± 3

139 ± 5

124 ± 4

117 ± 4

Seminal vesicles (mg)

470 ± 15

446 ± 21

357 ± 15

306 ± 14*

Prostate (mg)

49 ± 3

45 ± 4

30 ± 4

33 ± 4

Sperm parameters

Sperm concentration (106sperm/g caudal tissue)

633 ± 34

668 ± 32

1054 ± 52

1041 ± 60

Motile sperm (%)

65 ± 4

69 ± 5

55 ± 3

60 ± 1

Abnormal sperm (%)

2.1 ± 0.2

2.6 ± 0.3

4.5 ± 0.4

5.9 ± 1.3

aMean ±SE

bOrgan weights adjusted for bodyweight by analysis of covariance

* Significantly different from control (P < 0.05)

 

Table 3: Bodyweights and liver weights of female mice from the cross mating study and the offspring fertility test.

Observationa,b

Cross Mating Study

Offspring Fertility Test

Control

2.5 % EGPE in feed

Control

1.25% EGPE in feed

Number of females

38

17

18

16

Bodyweight (g)

37.1 ± 0.07

36.1 ± 0.9

30.97 ± 0.52

28.91 ± 0.50*

Liver (g)

2.15 ± 0.07

3.44 ± 0.11*

1.81 ± 0.04

2.08 ± 0.05*

aMean ±SE

bOrgan weights adjusted for bodyweight by analysis of covariance

* Significantly different from control (P < 0.05)

 

Table 4: Bodyweights of the offspring from the continuous breeding test (litter 5) at birth, weaning and the beginning of the offspring fertility test

Age (days)

Continuous Breeding Test

Control

0.25% EGPE in diet

1.25% EGPE in diet

2.5% EGPE in diet

Birth (Day 0)a

Male

1.77 ± 0.03 (34)

1.70 ± 0.04 (15)

1.69 ± 0.07 (18)

1.60 ± 0.04 (12)*

Female

1.68 ± 0.03 (34)

1.66 ± 0.05 (15)

1.66 ± 0.07 (18)

1.52 ± 0.05 (12)*

Weaning (Day 21)b

Male

9.82 ± 0.44 (41)

9.43 ± 0.45 (42)

7.35 ± 0.27 (33)*

5.84 ± 0.22 (32)*

Female

9.80 ± 0.41 (39)

8.34 ± 0.32 (54)

7.56 ± 0.29 (32)*

5.57 ± 0.24 (24)*

Mating trial (Day 74 ± 10)c

Male

35.57 ± 0.87 (20)

34.77 ± 0.70 (20)

30.82 ± 0.62 (20)*

28.76 ± 1.94 (3)*

Female

28.00 ± 0.62 (20)

27.15 ± 0.60 (20)

25.69 ± 0.37 (20)*

24.24 ± 0.82 (3)*

* Significantly different from control group (P < 0.05)

aMean ± SE for male or female pup weight per litter; number of litters evaluated indicated in parentheses

bMean ± SE for individual pup weights; number of pups weighed indicated in parentheses

cMean ± SE for individual mice

Conclusions:
Under the conditions of the test, administration of the test material did not affect the ability to produce 5 consecutive litters in the continuous breeding phase of the study; there was a small decrease in the number of pups/litter and in pup weight in the high dose group, 2.5%. When examined in a crossover study, the affected reproductive component was considered to be from the female, however this was considered inconclusive after assessment, with the only finding being reduced pup weight. Fertility was only minimally compromised, however neonatal toxicity was observed, with increased lethality observed throughout lactation, weaning and puberty. Second generation fertility was found not to be affected, a small decrease in live pup weight was observed. The test material was found to be toxic to immature mice of both sexes. Under the conditions of the test, the NOAEL for both parental and reproductive toxicity was found to be 400 mg/kg bw/day.
Executive summary:

The reproductive toxicity of the test material was investigated in Swiss CD-1 mice. Mice were exposed to the test material in diet at concentrations of 0, 0.25, 1.25 and 2.5% equivalent to 0, 0.4, 2.0 and 4 g/kg bw/day (a range finding study was conducted with concentrations of 0, 1.25 and 2.5, 5, 7.5 and 10% in feed for 2 weeks). Observations during the study included general toxicity, sperm analysis, reproductive performance, pup abnormalities and assessment of possible sex related effects (crossover mating).

Under the conditions of the test, administration of the test material did not affect the ability to produce 5 consecutive litters in the continuous breeding phase of the study; there was a small decrease in the number of pups/litter and in pup weight in the high dose group, 2.5%. When examined in a crossover study, the affected reproductive component was considered to be from the female, however this was considered inconclusive after assessment, with the only finding being reduced pup weight. Fertility was only minimally compromised, however neonatal toxicity was observed, with increased lethality observed throughout lactation, weaning and puberty. Second generation fertility was found not to be affected, a small decrease in live pup weight was observed. The test material was found to be toxic to immature mice of both sexes. Under the conditions of the test, the NOAEL for both parental and reproductive toxicity was found to be 400 mg/kg bw/day.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
400 mg/kg bw/day
Study duration:
chronic
Species:
mouse
Quality of whole database:
The quality of the database is high, the key study was performed in line with good scientific principles and reported in sufficient detail to address the endpoint. The lack of effects on fertility was confirmed in the supporting study.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

The key study, Heindel et al. (1990), studied the effects of 2-phenoxyethanol in Swiss CD-1 mice in a continuous breeding study. The study was performed in line with GLP according to the continuous breeding methodology which ensured that all stages of the reproductive cycle were encompassed in the test. Mice were exposed to the test material in diet at concentrations of 0, 0.25, 1.25 and 2.5% equivalent to 0, 0.4, 2.0 and 4 g/kg bw/day (a range finding study was conducted with concentrations of 0, 1.25 and 2.5, 5, 7.5 and 10% in feed for 2 weeks). Observations during the study included general toxicity, sperm analysis, reproductive performance, pup abnormalities and assessment of possible sex related effects (crossover mating). Under the conditions of the test, administration of the test material did not affect the ability to produce 5 consecutive litters in the continuous breeding phase of the study; there was a small decrease in the number of pups/litter and in pup weight in the high dose group, 2.5%. When examined in a crossover study, the affected reproductive component was considered to be from the female, however this was considered inconclusive after assessment, with the only finding being reduced pup weight. Fertility was only minimally compromised, however neonatal toxicity was observed, with increased lethality observed throughout lactation, weaning and puberty. Second generation fertility was found not to be affected, a small decrease in live pup weight was observed. The test material was found to be toxic to immature mice of both sexes. The study was reported to a high standard and was therefore assigned a reliability score of 2 in accordance with Klimisch (1997). Under the conditions of the test, the test material was found to have only minimal effects of fertility. A NOAEL of 400 mg/kg bw/day was concluded from the findings of the study.

The supporting study, Nagano et al. (1984), investigated the histopathological effects of 2-phenoxyethanol on the testes of male ICR mice. Male mice aged 6 weeks were dosed with the test material at 500 and 1000 mg/kg bw/day 5 days/week for 5 weeks via gastric intubation. Upon termination, testes were weighed and combined with the weight of the seminal vesicles and coagulating gland. Tissues were prepared for histopathological examination. Haematological changes were also investigated. The study was performed in line with good scientific standards, however, the results on 2-phenoxyethanol were reported in an assessment with several other chemicals using three different methods. The information presented on 2-phenoxyethanol was extremely brief, and therefore was assigned a reliability score of 3 in accordance with Klimisch (1997). Under the conditions of the test, the test material was found not to exhibit any effects on the testes of mice.

2-phenoxyethanol has been used as a read across substance since it is structurally similar to the substance to be registered. However it is anticipated to be more toxicologically active, and as such the results presented are taken to be the worst case scenario.


Short description of key information:
Oral feed: NOEL parental 400 mg/kg bw/day, NOEL 400 mg/kg bw/day, equivalent/similar to OECD 416, Heindel et al. (1990).

Justification for selection of Effect on fertility via oral route:
The continuous breeding study was selected as the key study as it was a thorough investigation on the effects of the fertility of the test material over multiple generations. The study was performed on a similar substance which was considered sufficiently similar to read-across the data to the registered substance in order to fulfil the data requirement.

Effects on developmental toxicity

Description of key information
Dermal: Maternal Toxicity NOEL 439 mg/kg bw/day, Embryotoxicity 71.4 mg/kg bw/day, equivalent/similar to OECD 414, Hoberman (1988).
Dermal: Maternal Toxicity NOEL 439 mg/kg bw/day, Embryotoxicity 143 mg/kg bw/day, equivalent/similar to OECD 414, Palmer et al. (1986).
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
71.4 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The overall quality of the database is good. Two studies on similar substance were available, both were assigned a reliability score of 2 in accordance with the principles for assessing data quality outlined in Klimisch (1997) as they were both reported to a high standard, and the methods followed were in basic compliance with the current standardised guidelines. The data is sufficient to address effects on development on the basis of weight of evidence.
Additional information

Palmer et al. (1986) investigated the developmental toxicity/teratogenicity of phenethyl alcohol, dosed dermally to the rat in a method similar to that of OECD 414. Undiluted phenethyl alcohol was administered to rats dermally at 0.07, 0.14, 0.28, 0.43 or 0.70 mL/kg/day from gestation day 6 through 15 at 24 hour intervals under an occluded dressing. Emphasis was placed on identifying whether vestigial cervical ribs in offspring were the most sensitive end point for developmental toxicity and whether they occurred in the absence of maternal toxicity. Signs of irritation were noted in all dams dosed with the test material, dose-dependently for onset, severity and duration. No maternal deaths occurred. Clinical signs of toxicity (ptosis and/or urine stained abdominal fur) were noted in the dams dosed with 0.70 mL.kg bw/day of the test material. Live foetal body weights were lower in litters from dams dosed with the test material, attaining statistical significance at doses of 0.14 mL/kg/day and above. Significant increase in the incidence of foetuses with cervical ribs occurred for the 0.70 mL/kg/day group. All other foetal observations were considered to be reversible delays in ossification. Delayed ossification (sternum and/or pelvic) was noted in the all test material administered groups. Delayed pelvic ossification was found to be significant in all groups dosed with the test material, but neither alteration demonstrated a clear dosage-dependent pattern. The study was performed in line with good scientific principles with a high level of detail in the reporting. The study was therefore assigned a reliability score of 2 in accordance with Klimisch (1997). Under the conditions of the study, on the basis of reduced fetal body weight, and ptosis and/or urine-stained abdominal fur in dams (and excluding irritation), the developmental and maternal no-effect levels for the test material were ≤0.07 and ≤0.43 mL/kg/day respectively.

 

Hoberman (1988) investigated the developmental toxicity effects of phenylethyl alcohol using cervical rib(s) as an indicator of teratogenicity. The study was performed to a method similar to OECD 414, however smaller group numbers were used (10 dams per dose level). Doses of 1.40, 0.43 and 0.14 mL/kg/day were administered occlusively to the skin to Crl rats between days 6 to 15 of gestation. Maternal animals were observed for signs of toxicity and local irritation. On gestation day 20, the animals were sacrificed and developmental parameters were assessed in the pups in utero. In addition to the developmental toxicity study, an absorption study was included to give a brief picture on the behaviour of the substance when dosed dermally. For this purpose 3 animals from the low and high dose groups received a radiolabelled dose on the final day of treatment. The test material was found to be rapidly and extensively absorbed in the pilot study. Both maternal and embryo-foetal toxicity was observed at a dose of 1.40 mL/kg bw/day (1428 mg/kg bw/day). The embryo-foetal toxic effects were not considered to be secondary to the maternal toxicity observed at this dose level. The 0.43 mL/kg bw/day (439 mg/kg bw/day) dose group was considered to be close to the threshold of maternal toxicity. A dose-dependent increase in the number of some morphological changes was recorded in the individual foetuses, litter values however were not affected. Treatment at 0.14 mL/kg bw/day (143 mg/kg bw/day) did not induce any maternal toxicity nor any adverse effects on litter values. Due to the changes noted in individual foetuses, in the 1428 and 439 mg/kg bw/day groups, the slight increases of morphological changes above the control values cannot be entirely discounted. The study was performed in line with good scientific principles and in accordance with the good laboratory practice. The level of reporting was good allowing sufficient assessment of the quality of the data. As such the study was assigned a reliability score of 2 in accordance with Klimisch (1997). Under the conditions of the test, the NOEL for maternal toxicity and embryotoxicity were 0.43 mL/kg bw/day (439 mg/kg bw/day) and 0.14 mL/kg bw/day (143 mg/kg bw/day), respectively.

Phenylethyl alcohol has been used as a read across substance since it is structurally similar to the substance to be registered. However it is expected to be more toxicologically active, and as such the results presented are taken to be the worst case scenario.


Justification for selection of Effect on developmental toxicity: via dermal route:
No single study selected since both studies are necessary to address effects on development through a weight of evidence approach.

Justification for classification or non-classification

According to the criteria outlined in Regulation (EC) No. 1272/2008 and Directive 67/548/EEC, the substance does not meet the criteria for classification for toxicity to reproduction.

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