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Administrative data

Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-study according to OECD guideline 416.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Principles of method if other than guideline:
not applicable
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
1-phenoxypropan-2-ol
EC Number:
212-222-7
EC Name:
1-phenoxypropan-2-ol
Cas Number:
770-35-4
Molecular formula:
C9H12O2
IUPAC Name:
1-phenoxypropan-2-ol
Constituent 2
Reference substance name:
1-phenylpropan-2-ol
EC Number:
211-821-0
EC Name:
1-phenylpropan-2-ol
Cas Number:
698-87-3
IUPAC Name:
1-phenylpropan-2-ol
Details on test material:
Identity: Protectol PP
Synonyms: Propylene Glycol Phenyl Ether, PPh
Purity: Isomeric mixture (85/15). Presumably, this means 85% 1-phenoxy-2-propanol (secondary alcohol, CAS No. 770-35-4) and 15% 2-phenoxy-1-propanol (primary alcohol, CAS No. 41593-38-8).

The stability of the test material in drinking water was established over a period of 96 hrs at room temperature and dose concentrations.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: not specified
- Age at study initiation: (P) x approximately 6 weeks; (F1) (continuous exposure)
- Weight at study initiation: (P) Males: 102 - 140 g; Females: 90 - 122g; (F1) Males: x-x g; Females: x-x g
- Fasting period before study: none
- Housing: individual
- Diet (e.g. ad libitum): not specified
- Water (e.g. ad libitum): not specified
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): standard conditions
- Humidity (%): standard conditions
- Air changes (per hr): standard conditions
- Photoperiod (hrs dark / hrs light): standard conditions

Administration / exposure

Route of administration:
oral: drinking water
Type of inhalation exposure (if applicable):
other: not applicable
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Propylene glycol phenyl ether was continuously administered to male & female F0 and F1 animals with drinking water at concentrations of 0, 100, 1000, 5000 ppm, respectively, until animals were terminated. Solutions were prepared once or twice a week. Propylene glycol phenyl ether concentrations were checked at start and at 3-monthly intervals during the administration period, and at its end

Details on mating procedure:
- M/F ratio per cage: 1:1 (male:female)
- Length of cohabitation: 3 weeks
- Proof of pregnancy: [sperm in vaginal smear] referred to as [day 0] of pregnancy
- For those pairs failing to mate, replacement of first male by another male with proven fertility was not done
- Further matings after two unsuccessful attempts: [no]
- After successful mating each pregnant female was caged (how): individual
- Any other deviations from standard protocol: not specified
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Propylene glycol phenyl ether concentrations were checked at start and at 3-monthly intervals during the administration period, and at its end. The stability of the test material in drinking water was established over a period of 96 hrs at room temperature and dose concentrations.
Duration of treatment / exposure:
Exposure period: 26 weeks
Premating exposure period (males): 77 days
Premating exposure period (females): 77 days
Duration of test: 40 weeks
Frequency of treatment:
7 days/week
Details on study schedule:
- F1 parental animals not mated until [approximately 10-13] weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were [21] days of age.
- Age at mating of the mated animals in the study: [10-13] weeks
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0 ppm
Basis:
nominal in water
Remarks:
Doses / Concentrations:
100 ppm (nominal in water)
Basis:
other: 11.3 mg/kg body weight/day (mean dose)
Remarks:
Doses / Concentrations:
1000 ppm (nominal in water)
Basis:
other: 113.9 mg/kg body weight/day (mean dose)
Remarks:
Doses / Concentrations:
5000 ppm (nominal in water)
Basis:
other: 477.5 mg/kg body weight/day (mean dose)
No. of animals per sex per dose:
25 rats/sex/dose
Control animals:
yes
Details on study design:
- Dose selection rationale: not specified
- Rationale for animal assignment: random
Positive control:
not applicable

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights of parental animals were recorded weekly with the following exceptions for females:
-during mating periods females were weighed on day 0 and on days 7, 14, 20 post coitum
-females without evidence of sperm were not weighed during the mating period
-females with litters were weighed on days 1,4,7,14 and 21 post partum
-females without litters were not weighed during lactation period
-F0 and F1 females were weighed after weaning of the last F1 or F2 pups, parallel to the F0 and F1 males, once weekly until termination

FOOD CONSUMPTION: Yes
- Time schedule for examinations: Food consumption was determined once a week during the premating phase of both F0 and F1, and weekly during
gestation. During lactation it was determined weekly on days 1, 4, 7 and 14, but not during 14-21 as required by the guideline since the pups start to consume considerable amounts of solid food. Food consumption was not determined for F0 and F1 males after the premating phase and for females without evidence of sperm or without litters in the lactation phase

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water consumption of F0 and F1 parental animals was determined once a week during the premating periods. After premating (10th week), water consumption of females during gestation was determined for days 0-1, 6-7, 13-14 and 19-20 post coitum, and during lactation period for days 1-2, 4-5, 7-8 and 14-15 pp (post parturition). After day 15 pp water consumption of the F0 and F1 dams was not determined since from then onwards the pups begin to consume considerable amounts of water. Water consumption was not determined for F0 and F1 males after the premating phase and for females without evidence of sperm or without litters in the lactation phase. Intake of PPh was calculated from the daily water consumption.

Oestrous cyclicity (parental animals):
Estrous cycle length and normality were evaluated for all F0 and F1 females for a minimum of 3 weeks prior to mating; this was continued throughout the mating period. Vaginal smear was examined at necropsy to determine the stage of the estrous cycle for each F0 and F1 female rats
Sperm parameters (parental animals):
Parameters examined in [all] male parental generations: Sperm parameters were determined (sperm motility, morphology, sperm head count in testis and in cauda epididymis) immediately after necropsy and weighing the right testis and cauda epididymis. Sperm motility examinations were randomized; sperm morphology and sperm head count were evaluated in control and highest dose animals only
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: [yes]
- If yes, maximum of [8] pups/litter ([4]/sex/litter as nearly as possible); excess pups were killed and discarded. Also, litters with less than 8 pups were not standardized

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring: [number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities]

GROSS EXAMINATION OF DEAD PUPS:
[yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead.]
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [after the last litter of each generation was weaned.]

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.] Organ weights of all F0 and F1 parental animals terminated at schedule were determined: body weight, liver, kidneys, adrenals, testes, epididymides (total, cauda), prostate
gland, seminal vesicles with coagulation glands, ovaries, uterus (with cervix and oviducts), thymus, spleen, brain, pituitary gland

HISTOPATHOLOGY / ORGAN WEIGHTS
- The following organs were fixed or embedded for histopathology: vagina, cervix uteri, uterus, ovaries, oviducts, left testicle and epididymides, seminal vesicles, coagulating glands, prostate gland, pituitary gland, liver, kidneys, urinary bladder, thymus, spleen, brain, adrenal glands, and all gross lesions. In ovaries, a Differential Ovarian Follicle Count (DOFC) was also included
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [21] days of age.
- These animals were subjected to postmortem examinations (macroscopic examination) as follows: all pups with scheduled termination, i.e. those culled on day 4 pp, and those terminated at day 21, were examined externally and eviscerated; organs were assessed macroscopically, and additionally, if this was deemed necessary due to notable findings and abnormalities. The same procedure was applied to all stillborn pups and all pups that died up to weaning. After scheduled termination, of the pups organs brain, spleen and thymus were weighed of one pup/sex and litter from F1 and F2 pups.

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS
- not specified
Statistics:
Extensive statistical evaluation of the clinical data included the use of the Dunnett-test for comparison with the control group, Fisher's exact test, Wilcoxon test and Kruskal-Wallis-test. Statistical evaluation of the organ weight parameters involved Kruskal-Wallis test and Wilcoxon test, if p was equal to or < 0.05. Follicle data from DOFC were evaluated using a Wilcoxon test
Reproductive indices:
Male reproduction indices (mating and fertility index) were calculated. For females, indices pertaining to mating, fertility, gestation were calculated.
Offspring viability indices:
For F1 and F2 litters, live birth index was calculated (percentage of liveborn pups). Postimplantation loss was calculated after termination of females from the number of implantations and pups delivered

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS) - No substance-related adverse effects seen with respect to clinical examination.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS) - F0 animals - In the 5000 ppm group, compared with controls significant reductions in consumption of water during premating (-20% males, -22% females), gestation (-21%) and lactation (-19%) and in food consumption (significant on some days) during premating (-6% males, -5% females), gestation (-5%) and lactation (-8%). This was paralleled by a clearly decreased body weight (bw) and body weight change (bwc) in males (-10% each). In females, reduced values were also seen during premating, gestation and lactation (bw: -6, -7, -11%; bwc: -8, -14, -8%)
F1 animals - Significantly decreased water consumption during premating (up to -20% males, -29% females), gestation (-25%) and lactation (-33%). Mainly significantly reduced food consumption during premating (-8% males and females), gestation (-12%) and lactation (-18%). Clearly decreased bw and bwc in males (bw -19%, bwc ca. -10%). In females, reduced values were seen during premating, gestation and lactation (bw: -22, -15, -16%; bwc: +/-0, -25, -31%). Though clear effects on body weight were seen in both, males and females, level of significance was achieved only for single periods

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS) - F0 animals - In the 5000 ppm group, compared with controls significant reductions in consumption of water during premating (-20% males, -22% females), gestation (-21%) and lactation (-19%) and in food consumption (significant on some days) during premating (-6% males, -5% females), gestation (-5%) and lactation (-8%). This was paralleled by a clearly decreased body weight (bw) and body weight change (bwc) in males (-10% each). In females, reduced values were also seen during premating, gestation and lactation (bw: -6, -7, -11%; bwc: -8, -14, -8%)
F1 animals - Significantly decreased water consumption during premating (up to -20% males, -29% females), gestation (-25%) and lactation (-33%). Mainly significantly reduced food consumption during premating (-8% males and females), gestation (-12%) and lactation (-18%). Clearly decreased bw and bwc in males (bw -19%, bwc ca. -10%). In females, reduced values were seen during premating, gestation and lactation (bw: -22, -15, -16%; bwc: +/-0, -25, -31%). Though clear effects on body weight were seen in both, males and females, level of significance was achieved only for single periods

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS) - No substance-related adverse effects seen with respect to reproductive function

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS) - No substance-related adverse effects seen with respect to reproductive function

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS) - No substance-related adverse effects seen with respect to reproductive performance

ORGAN WEIGHTS (PARENTAL ANIMALS) -No substance-related adverse effects seen with respect to organ weights

GROSS PATHOLOGY (PARENTAL ANIMALS) -No substance-related adverse effects seen with respect to pathology

HISTOPATHOLOGY (PARENTAL ANIMALS) - No substance-related adverse effects seen with respect to histopathology

OTHER FINDINGS - Propylene glycol phenyl ether intake by premating F0 and F1 females was slightly enhanced compared with values for males (total treatment period) in all dose groups. PPh intake was markedly enhanced during gestation and lactation when compared with premating animals (up to ca. 1.5 fold in F0 females). Intake of PPh was also enhanced in F1 parental animals compared with F0 parents

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
5 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: based on reproductive performance and fertility
Dose descriptor:
NOAEL
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Details on results (F1)

VIABILITY (OFFSPRING) - No substance-related adverse effects seen with respect to viability of the offspring

CLINICAL SIGNS (OFFSPRING) - No substance-related adverse effects seen with respect to clinical examination, sexual maturation (F1 pups only), pup organ weights, pathology

BODY WEIGHT (OFFSPRING) - F1 pups - Significantly lower bw at weaning on d 21 (-11%, both sexes combined) and lower bwc (-13%). Concomitantly a retardation in sexual maturation as evidenced by delayed preputial separation and vaginal opening in the selected F1 male and female animals
F2 pups - Significantly lower mean bw in male pups from day 4 pp (post parturition) onwards. During d 7-21 pp lowered by 23% (both sexes combined). Significantly impaired body weight change in male and female pups (-26%; d 4-21 pp)

SEXUAL MATURATION (OFFSPRING) - F1 pups - Significantly lower bw at weaning on d 21 (-11%, both sexes combined) and lower bwc (-13%). Concomitantly a retardation in sexual maturation as evidenced by delayed preputial separation and vaginal opening in the selected F1 male and female animals

ORGAN WEIGHTS (OFFSPRING) - F2 pups - During pathological examinations, organ weight changes (both sexes combined) were seen as follows: significantly lower mean absolute weights of brain (-6%), thymus (-23%) and spleen (-34%) compared to controls. Significant relative organ weight changes were noted for brain (+30%) and spleen (-18%). The relationship to treatment of reduced relative spleen weights to body weights in the progeny of high dose animals was unclear, since a similar effect was not noted in the parental animals

GROSS PATHOLOGY (OFFSPRING) - No substance-related adverse effects seen with respect to clinical examination, sexual maturation (F1 pups only), pup organ weights, pathology

OTHER FINDINGS - Propylene glycol phenyl ether intake by premating F0 and F1 females was slightly enhanced compared with values for males (total treatment period) in all dose groups. PPh intake was markedly enhanced during gestation and lactation when compared with premating animals (up to ca. 1.5 fold in F0 females). Intake of PPh was also enhanced in F1 parental animals compared with F0 parents

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Results: F2 generation

Effect levels (F2)

Dose descriptor:
NOAEL
Generation:
F2
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

none

Applicant's summary and conclusion

Conclusions:
Propylene glycol phenyl ether was continuously administered with drinking water to rats over two parental generations at concentrations of 0, 100, 1000 and 5000 ppm. Under the conditions of this study, NOAELs were established as follows: NOAEL for reproductive performance and fertility: 5000 ppm (about 477.5 mg PPh/kg body weight/day) for the F0 and F1 parents; NOAEL for developmental toxicity: 1000 ppm (about 113.9 mg PPh/kg body weight/day) for the F1 and F2 progeny; NOAEL for general systemic toxicity: 1000 ppm (about 113.9 mg PPh/kg body weight/day) for the F0 and F1 parents. Thus, developmental toxicity was seen only at a dose which was also toxic to the parent animals. No sign of teratogenicity was seen at either dose in this study.

Reproductive performance or fertility was not affected in F0 or F1 parental animals of either dose group. Estrous cycle, mating behavior, conception, gestation, parturition, lactation and weaning as well as sperm parameters, sexual organ weights, and gross and histopathological findings of these organs were similar between control and treated animals.

Signs of general, systemic toxicity were noted in both parental generations (F0 and F1) in groups receiving 5000 ppm, but not in others. Toxicity was characterized by decreased water and food consumption, decreased body weight and body weight gain in parental F0 an F1 males and females. Pathology and histopathology did not reveal substance-related adverse effects in F0 and F1 parental animals.

The clinical, gross and histopathological examinations in F0 and F1 parental animals from the low and intermediate dose groups did not yield any indication of systemic toxicity.

Substance-related signs of developmental toxicity were seen in progeny of the high dose (5000 ppm) F0 and F1 parents in terms of reduced pup body weight and body weight gain. This is directly related to lower absolute weights of the thymus, spleen and brain in pups and delayed sexual maturation. Moreover, reproduction parameters of these animals were not adversely affected after gaining sexual maturity. This supports the view that delayed preputial separation and vaginal opening resulted from a general retardation of physical development. No signs of developmental toxicity were seen in pups from groups receiving medium or low doses (1000 or 100 ppm, respectively.).

Under the conditions of this study, NOAELs were established as follows:
NOAEL for reproductive performance and fertility: 5000 ppm (about 477.5 mg PPh/kg body weight/day) for the F0 and F1 parents
NOAEL for developmental toxicity: 1000 ppm (about 113.9 mg PPh/kg body weight/day) for the F1 and F2 progeny
NOAEL for general systemic toxicity: 1000 ppm (about 113.9 mg PPh/kg body weight/day) for the F0 and F1 parents

Thus, developmental toxicity was seen only at a dose which was also toxic to the parent animals. No sign of teratogenicity was seen at either dose in this study.
Executive summary:

In this study, Propylene glycol phenyl ether was continuously administered with drinking water to rats over two parental generations at concentrations of 0, 100, 1000 and 5000 ppm.

Reproductive performance or fertility was not affected in F0 or F1 parental animals of either dose group. Estrous cycle, mating behavior, conception, gestation, parturition, lactation and weaning as well as sperm parameters, sexual organ weights, and gross and histopathological findings of these organs were similar between control and treated animals.

Signs of general, systemic toxicity were noted in both parental generations (F0 and F1) in groups receiving 5000 ppm, but not in others. Toxicity was characterized by decreased water and food consumption, decreased body weight and body weight gain in parental F0 an F1 males and females. Pathology and histopathology did not reveal substance-related adverse effects in F0 and F1 parental animals.

The clinical, gross and histopathological examinations in F0 and F1 parental animals from the low and intermediate dose groups did not yield any indication of systemic toxicity.

Substance-related signs of developmental toxicity were seen in progeny of the high dose (5000 ppm) F0 and F1 parents in terms of reduced pup body weight and body weight gain. This is directly related to lower absolute weights of the thymus, spleen and brain in pups and delayed sexual maturation. Moreover, reproduction parameters of these animals were not adversely affected after gaining sexual maturity. This supports the view that delayed preputial separation and vaginal opening resulted from a general retardation of physical development. No signs of developmental toxicity were seen in pups from groups receiving medium or low doses (1000 or 100 ppm, respectively.).

Under the conditions of this study, NOAELs were established as follows:

NOAEL for reproductive performance and fertility: 5000 ppm (about 477.5 mg PPh/kg body weight/day) for the F0 and F1 parents

NOAEL for developmental toxicity: 1000 ppm (about 113.9 mg PPh/kg body weight/day) for the F1 and F2 progeny

NOAEL for general systemic toxicity: 1000 ppm (about 113.9 mg PPh/kg body weight/day) for the F0 and F1 parents

Thus, developmental toxicity was seen only at a dose which was also toxic to the parent animals. No sign of teratogenicity was seen at either dose in this study.