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Key value for chemical safety assessment

Effects on fertility

Description of key information
OECD 416, rat, 2 gen, oral: not toxic to reproduction
NOAELrep = 300 mg/kg bw/day; LOAELrep > 300 mg/kg bw/day
Link to relevant study records
Reference
Endpoint:
two-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
According to Guideline.
TEST ANIMALS
- Source: Charles River UK Ltd., England
- Age at study initiation: 4-5 weeks
- Weight at order: 120 - 150 g
- Age after acclimatisation: males 6-7 weeks, females 14-15 weeks: Mating of F1 animals started at an age of 16 weeks
- Fasting period before study: No
- Housing: Group housed, except for mating period. During pregnancy and lactation period females were housed individually.
- Diet (ad libitum): Pelleted SQC rat and mouse No. 3 Breeder
- Water (ad libitum): Purified water
- Acclimation period: males 12 days, females 6 - 7 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 27
- Humidity (%): 36 - 62
- Air changes (per hr): 16
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: drinking water
Vehicle:
water
Details on exposure:
The test article was formulated for dosing as solutions in the vehicle, purified drinking water. For formulation, the weighed quantity of test article was mixed with the appropriate volume of vehicle. Separate formulations were prepared for each dose level. Formulations were prepared once weekly and stored in polycarbonate aspirators at ambient temperature in the animal room during the week of use.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: Maximum 14 days
- Proof of pregnancy: Sperm in vaginal smear referred to as day 0 of pregnancy
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: No
- After successful mating each pregnant female was caged: Individually
- Any other deviations from standard protocol: Due to reduced numbers of litters in each group of the F1a generation the F0 generation was re-paired following weaning of the F1a generation to produce a F1b generation. Females were allowed to rear their offspring to weaning on day 21 post partum.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Two 10 mL samples from the middle of the aspirator of each dose group prepared for weeks 1, 5, 9, 13, 17, 21, 25, 29 and 33 have been quantified via HPLC analysis using an internal standard for quantification. The actual concentrations proved the nominal concentrations.
Duration of treatment / exposure:
P males: 11 weeks before mating, during both mating periods (for F1a and F1b), until necropsy (total 198 days).
P females: 2 weeks before mating, during both mating periods (for F1a and F1b), pregnancy and lactation, until necropsy (Day 21 post partum) (total 71 days for F0 to F1a and 114 days for F0 to F1b).
F1 males were dosed from birth until approximately 16 weeks of age, during mating, until necropsy (total 110 days).
F1 females were dosed from birth until approximately 16 weeks of age, during mating, pregnancy and lactation period (total 119 days).
Frequency of treatment:
Daily
Details on study schedule:
F1 parental animals were mated 16 weeks after selected from the F1 litters.
Dose / conc.:
30 other: mg/kg bw/day, calculated by test article intake (rough estimate, actual values are rather higher)
Dose / conc.:
100 other: mg/kg bw/day, calculated by test article intake (rough estimate, actual values are rather higher)
Dose / conc.:
300 other: mg/kg bw/day, calculated by test article intake (rough estimate, actual values are rather higher)
Dose / conc.:
0.03 other: % in water, concentration proven by analytcial examination
Dose / conc.:
0.1 other: % in water, concentration proven by analytcial examination
Dose / conc.:
0.3 other: % in water, concentration proven by analytcial examination
No. of animals per sex per dose:
30 (F0 gen), 25 (F1 gen)
Control animals:
yes, concurrent vehicle
Positive control:
Not required.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
Twice daily for mortalities and once daily for clinical signs of toxicity

BODY WEIGHT: Yes
Males: at weekly intervals. Females weekly during pre-mating, on days 0, 7, 14 and 20 of pregnancy and on days 0, 4, 7, 14, 21 post partum.

FOOD CONSUMPTION: Yes
At weekly intervals. Additionally for females from days 1-4, 4-7, 7-11 and 11-14 post partum

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
Daily for males and females during pre-mating, for males at the end of the mating period and for females during pregnancy and lactation. Water consumption was not measured for mating pairs.

Other examinations:
Blood samples were taken from 10 males and 10 females of each dose group one week prior the F0-F1a and the F1-F2 mating period. Following parameter were assessed: leucocyte differential count, total leucocyte count, A/G ratio, alanine aminotransferase, albumin, alkaline phosphatase, aspartate aminotransferase, blood urea nitrogen, creatinine, globulin, total bilirubin, total protein, triglycerides
Oestrous cyclicity (parental animals):
Beginning two weeks before the start of the first F0 and the F1 mating period and during all the mating periods until confirmation of maing or end of the relevant periods vaginal smears were taken daily and examined for estrous cycle stage.
Sperm parameters (parental animals):
Parameters examined in male parental generations:
testis weight, epididymis weight, sperm count in epididymides, percent motile sperms (incl. calculation of straight line velocity and average pathe velocity), sperm motility, sperm morphology
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.

PARAMETERS EXAMINED
The following parameters were examined in F1 and F2 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, body weight and body weight gain, physical or behavioural abnormalities, time until developmental milestones are reached (e.g. ear opening, balanopreputial separation of males, vaginal perforation of femals, etc.)

FUNCTIONAL OBSERVATIONS
On day 22 post partum the auditory startle habituation response was evaluated using the SR-Screening System on pups selected for special necropsy. At 28 days of age the selected pups were examined for learning potential using a water filled e-maze in two session on consecutive days. A session consisted of six runs. At approximately 28-35 days post partum the selected F1-generation pups were tested for motor activity.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead. Necropsy was performed on all pups killed prematurely or found dead and for surplus pups after selection for special necropsy and rearing
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals after the mating period
- 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
Following organs were weighed: brain, pituitary gland, liver, adrenals, spleen, thymus, kidneys, testes, epididymides (toatl and cauda), seminal vesicles, prostate gland, coagulating gland, ovaries, oviducts, uterus, cervix, vagina

HISTOPATHOLOGY
The following tissues were prepared for microscopic examination: adrenals, aorta, brain, caecum, colon, duodenum, epididymides, femur, ileum, jejunum, kidneys, liver, mesenteric lymph node, oesophagus, ovaries, pituitary, prostate, rectum, sciatic nerve, seminal vesicles, spinal cord, spleen, stomach, submandibular lymph node, testes, thymus, thyroids, uterus, vagina, gross lessions.
Detailed evaluation were further performed on opened stomach to further examine the glandular and non-glandular regions of the stomach for possible signs of irritation.
Postmortem examinations (offspring):
SACRIFICE
Necropsy was conducted on all pups killed prematurely or found dead and for surplus pups after selection for special necropsy and rearing.

GROSS NECROPSY
- One male and one female pup from each of the first twenty litters comprising sufficient pups were selected from F1a and F2 litters for special necropsy. Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera. Special necropsy comprised assessment of organ weights and fixation in neutral buffered formaldehyde.

ORGAN WEIGTHS
The following tissues were weighed during special necropsy: brain, liver, adrenals, spleen, thymus, kidneys, testes, ovaries.

HISTOPATHOLOGY
The following tissues were prepared for microscopic examination of the F1 generation: adrenals, aorta, brain, caecum, colon, duodenum, epididymides, femur, ileum, jejunum, kidneys, liver, mesenteric lymph node, oesophagus, ovaries, pituitary, prostate, rectum, sciatic nerve, seminal vesicles, spinal cord, spleen, stomach, submandibular lymph node, testes, thymus, thyroids, uterus, vagina, gross lessions.
Statistics:
Group means and standard deviations were calculated for each observation time. Analysis of variance (ANOVA) was performed on all parameters. Heterogeneity of variance was analysed using Levenes test. Williams test was performed to compare high dose with control at the two-sided 5% level. Kruskal-Wallis ANOVA was performed to assess overall differences between treatment groups followed by Shirley´s test.
Reproductive indices:
Fertility index, gestation index, copulation index, sex ratio
Offspring viability indices:
live birth index, viability indices
Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY:
No test item related clinical signs of toxicity were observed. 2, 2, 3 and 5 females were found dead or were killed prematurely during the F0 generation phase at 0, 30, 100 and 300 mg/kg bw/d. The observation and timings of these losses do not support any relationship to dosage and mortalities are considered to be not treatment related.

BODYWEIGHT:
(Transient) reductions in F0 males and females during the first two weeks of the study were considered to be related to palatability issues.

BIOCHEMISTRY:
Reduced triglyceride levels in F0 females was observed. The values were however within the historical control.

SPERM MEASURES:
Slight but significantly reduced straight line velocity (VSL) of the sperm was observed at the top dose (19.5 ± 8.8 compared to 25.1 ± 10.9 µm/s) without any significant effects on averaged path velocity (VAP) or total motility. No effect on sperm morphology was observed.

BODY WEIGHT AND WATER CONSUMPTION (PARENTAL ANIMALS)
No test item related effect on body weight was observed. The water consumption at the top dose was lower compared to the other groups. This is most likely due to palatability problems.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
No test item related effects observed.

ORGAN WEIGHTS:
The male F0 generation showed a small but significant reduction in body weight-liver weight ratios, but the corresponding brain related liver weights and the absolute liver weights developed not in a dose dependant way. For the F1 generation where similar results were reported, no dose-response relationship was detected either. No influence on liver weight development was seen in the F2 generation. None of the groups revealed any histopathological or clinical-chemical findings, which could be attributed to hepatotoxicity. This led to the conclusion that this untypical liver weight reduction was of no toxicological relevance.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No test item related effects on the copulation index was observed. The fertility index was low in each group of the F0 generation. Consequently F0 females were paired after weaning of F1a generation with a different male from the original pairing. The fertility index at 300 mg/kg bw/d was slightly decreased compare to the other groups for F0-F1b females. No effect on fertility was observed in the F1-F2 generation. Thus this finding was considered to be not treatment related. No treatment related effect on duration of gestation and gestation index were observed.

GROSS PATHOLOGY (PARENTAL ANIMALS)
No test item related effects

HISTOPATHOLOGY (PARENTAL ANIMALS)
No test item related effects were observed, except for slight signs of local irritation in the stomach at the top dose.
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
300 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No treatment-related effects observed.
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
300 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No treatment-related effects observed.
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
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
CLINICAL SIGNS AND MORTALITY:
No test item related clinical signs of toxicity or mortality was observed. At the mid dose of the F1a generation a large portion of pups showing poor or no maternal care were observed. 36% of those pups from 6 litters died by day 2 post partum. The same finding was observed at control and low dose pups of the F2 generation. These findings showed no does response and were considered to be not treatment related.

BODYWEIGHT:
A slight not significant reduction in body weight and body weight gain for males and female pups of the F1a generation at the top dose was observed. As body weight and body weight gain was slightly and not significantly higher in the F1b and the F2 generation this finding was considered to be not treatment related.

BIOCHEMISTRY:
An increased percentage neutrophil count in F1 males was observed. This finding was not observed in F0 males and was within the historical control. Thus this finding was not treatment related.

SEXUAL MATURATION:
A significant increase in time taken for sexual development of females at the top dose was observed. For males a not significant increase in time taken for sexual maturation was observed at the top dose. No difference in body weight of these females was observed. No effect on fertility or mating performance was observed.

ORGAN WEIGHTS:
see 'parental animals'

VIABILITY (OFFSPRING): No effects on pup survival (day 4 - 21 post partum), live birth index and mean litter size were observed. Pup survival of the F1a generation (day 0 - 4 post partum) was slightly lower in the treated groups when compared to control. The differences were not significant, showed no does reponse relationship and were not observed for F1b and F2 pups. Therefore this finding was considered to be not treatment related.

GROSS PATHOLOGY (OFFSPRING)
Pups showing a lack of maternal care and which died or were killed moribundly showed signs of canibalisation and had no milk in the stomach.
No treatment related effects were observed.

SPECIAL NECROPSY
No treatment related macroscopic findings were observed.

HISTOPATHOLOGY (OFFSPRING)
No effects were observed.

OTHER FINDINGS (OFFSPRING)
No adverse treatment related effect on the time to developmental milestones was observed. No treatment related effects on learing ability and functional observational battery were observed.
Dose descriptor:
NOAEL
Remarks:
systemic
Generation:
F1
Effect level:
300 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No treatment-related effects observed.
Dose descriptor:
NOAEL
Remarks:
reproduction
Generation:
F1
Effect level:
300 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No treatment-related effects observed.
Reproductive effects observed:
no

Slight but significantly reduced straight line velocity (VSL) of the sperm was without any significant effects on averaged path velocity (VAP) or total motility. Moreover, in the available subchronic and chronic toxicity studies on various AES the primary sex organs of the males and females did not show evidence for treatment-related adverse effects.The observed reduced triglyceride levels (female) and increased percentage neutrophil counts (males) were slight and within the range of the historical control data.

The male F0 generation showed a small but significant reduction in bodyweight-liver weight ratios, but the corresponding brain related liver weights and the absolute liver weights developed not in a dose dependant way. For the F1 generation where similar results were reported, no dose-response relationship was detected either. No influence on liver weight development was seen in the F2 generation. None of the groups revealed any histopathological or clinical-chemical findings, which could be attributed to hepatotoxicity. This led to the conclusion that this untypical liver weight reduction was of no toxicological relevance, additionally underlined by the absence of such effects in the studies for subchronic toxicity mentioned above.

There was evidence of toxicity on pup development at the top dose that was characterised by an increase in the time taken for sexual development of the male (not significant) and female (significant) offspring.This was investigated in more detail in the developmental toxicity study up to 1.5 g/kg bw and no effects were noted there. Considering all these facts the subchronic NOAEL for systemic toxicity and reproduction toxicity can be set to greater than 300 mg/kg bw.

Conclusions:
In summary, there was no effect of treatment at any dose level on reproduction of the parents or offspring (NOAEL >= 0.3 %; > 300 mg/kg/day).
Based on this study an overall NOAEL for systemic effects of 0.3 % (300 mg/kg bw) can be deduced.
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The whole data base is conclusive and of high quality.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

One study on reproductive toxicity is available for AES (C12-14) Na (CAS 68891-38-3). In a two-generation reproduction study with AES (C12-14) Na (CAS 68891-38-3, 27% a.i.)(BASF, 1999) Sprague Dawley rats were dosed via drinking water at 0, 0.03, 0.1 and 0.3%, which corresponded to daily doses of ca. 0, 30, 100 and 300 mg/kg bw/day. There were some changes indicative of parental toxicity in the group treated with 0.3% of the test substance. Slight but significantly reduced straight line velocity (VSL) of the sperm was without any significant effects on averaged path velocity (VAP) or total motility. Moreover, in the available subchronic and chronic toxicity studies on various AES the primary sex organs of the males and females did not show evidence for treatment-related adverse effects. The observed reduced triglyceride levels (female) and increased percentage neutrophil counts (males) were slight and within the range of the historical control data. The male F0 generation showed a small but significant reduction in body weight-liver weight ratios, but the corresponding brain related liver weights and the absolute liver weights developed not in a dose dependent way. For the F1 generation where similar results were reported, no dose-response relationship was detected either. No influence on liver weight development was seen in the F2 generation. None of the groups revealed any histopathological or clinical-chemical findings, which could be attributed to hepatotoxicity. This led to the conclusion that this untypical liver weight reduction was of no toxicological relevance, additionally underlined by the absence of such effects in the studies for subchronic toxicity mentioned above. There was evidence of toxicity on pup development at this dose level that was characterised by an increase in the time taken for sexual development of the male (not significant) and female (significant) offspring. This was investigated in more detail in the developmental toxicity study up to 1500 mg/kg bw/day (see "Developmental toxicity") and no effects were noted there. Considering all these facts, the subchronic NOAEL for systemic toxicity can be set to greater than 300 mg/kg bw/day.

Effects on developmental toxicity

Description of key information

OECD 414, rat, developmental toxicity, oral: not teratogenic

NOAELdev = 1000 mg/kg bw/day; LOAELdev > 1000 mg/kg bw/day

OECD 414, rabbit, developmental toxicity, oral: not teratogenic

NOAELdev = 300 mg/kg bw/day; LOAELdev > 300 mg/kg bw/day

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
8 June - 15 July 1993
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Administration comprised only period of organogenesis.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Remarks:
CD
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Wiga Sulzfeld, Germany
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: approx. 204 g
- Number of animals: 96, primiparous time-mated females
- Housing: Individually in Makrolon Type M3 cage (Ebeco, 44579 Castrop-Rauxel, Germany) with standard softwood bedding (ARWI-Center, 45307 Essen, Germany)
- Diet: Pelleted Altromin Maintenance Diet 1324, ad libitum
- Water: Community tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 23
- Humidity (%): 42 - 66
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 8 June 1993 To: 15 July 1993
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing solutions were prepared daily by dissolving appropriate amounts of the test material in water yielding a final concentration of 1, 3, and 10% corresponding to 100, 300 and 1000 mg/kg bw/day, respectively.

VEHICLE:
- Concentration in vehicle: 1, 3 and 10%
- Amount of vehicle: 10 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The formulation was analysed by "Henkel TTA-Zentrale Analytik" in the time from 21 - 30 June 1993 (study number TTA 93-6431). The concentrations of the test substance in aqua dest. based upon the results of the determination of the dry sediment confirmed the active content of 70.1% of the test substance.
Details on mating procedure:
- Impregnation procedure: purchased timed pregnant
The females were mated at the supplier with an accurate day of mating. They were received at the testing facility on day 0 of gestation.
The mated animals (number) were delivered:
June 08, 1993 (24)*
June 09, 1993 (24)
June 23, 1993 (24)
June 24, 1993 (18)
June 25, 1993 (16)**
*: on delivery 3 animals dead, 1 animal died one day later
**: 15 females used in study
Duration of treatment / exposure:
Day 6-15 (incl.), post coitum
Frequency of treatment:
daily, 7 days/week
Duration of test:
Until Day 20 post coitum
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Remarks:
corrected for a.i. content of test material
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Remarks:
corrected for a.i. content of test material
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Remarks:
corrected for a.i. content of test material
No. of animals per sex per dose:
24 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The dose selection is based on the results of an earlier toxicoligical investigation. Rats (10 animals/dose/sex) were orally exposed to 100, 500 and 1000 mg/kg bw/day for 28 days (BASF, 1985, TBD870334). Adverse effects (lesions in the mucosa of the forestomach) were observed at 500 and 1000 mg/kg bw/day. In addition, alterations in hematology and clinical chemistry parameters were observed in the mid- and high-dose group animals when compared to the controls. Therefore, 100, 300 and 1000 mg/kg bw/day were selected as the dose levels for the pre-natal developmental toxicity study since the females in this study were dosed on only 10 consecutive days (Day 6 - 15, inclsive, post coitum).
Maternal examinations:
CAGE SIDE OBSERVATIONS AND MORTALITY: Yes
- Time schedule: twice daily
- Cage side observations included: mortality, signs of reaction to treatment and symptoms of illness

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 6, 16 and 20 post coitum

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on Day 20 post coitum by an overdose of ether
- Organs examined: all maternal organs
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes

Examinations included: Gravid uterus weight, number of corpora lutea, implantations and early/late resorptions


The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes, all per litter
- Soft tissue examinations: Yes, half per litter
- Skeletal examinations: Yes, half per litter
- Head examinations: No
Statistics:
If the variables could be assumed to follow a normal distribution, the Dunnett-Test, based on a pooled variance, was applied for the comparison between the treated groups and the control group.
The Steel-Test was applied when the data could not be assumed to follow a normal distribution.
Fisher's Exact test for 2x2 tables was applied if the variables could be dichotomised without loss of information (Bonferroni-Holm-corrected).
Clinical signs:
no effects observed
Description (incidence and severity):
No test substance-related symptoms were observed in any animal at any dose level.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
No deaths occured in any dams of the control and treatment groups.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weights and body weight gains were essentially similar in all groups and comparable with the controls.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No macroscopic changes were noted in the dams of the control and treatment groups.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
In the mid-dose group (300 mg/kg bw/day), the mean value of the pre-implantation loss was decreased (level 5%). The finding was considered incidental and in the normal range.
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Remarks:
corrected for a.i. content of test material
Basis for effect level:
other: No treatment-related effects observed.
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
effects observed, non-treatment-related
Description (incidence and severity):
In the high-dose group (1000 mg/kg bw/day), the mean value of the total males was increased (level 5%) and that of the total females decreased (level 5%). These findings were considered to be incidental and in the normal range.
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
The statistically significant differences were considered to be incidental. The finding "two sternebrae, non ossified" is not a negative effect of the substance because a decrease indicate a more developed embryonic stage than in the other fetuses. The retardation effect of the "hyoid, incomplete ossified" is also incidental and not dose-related. Only singular litters were affected. The incidental character of this variation is emphasised by the fact the values were within the normal range of variation of this strain.
Visceral malformations:
no effects observed
Other effects:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Remarks:
corrected for a.i. content of test material
Sex:
male/female
Basis for effect level:
other: No treatment-related effects observed.
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Conclusions:
The results of this study showed that repeated oral administration (Day 6-15 post coitum) of the test substance to pregnant rats caused no symptoms of cumulative toxicity and does not reveal any embryotoxic or teratogenic potential up to a dose level of 1000 mg/kg body weight/day.
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose for cross-reference:
read-across source
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
>= 300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: No relevant treatment-related effects observed.
Remarks on result:
other: Source, key, 68585-34-2, 1972
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
>= 165 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: No relevant treatment-related effects observed.
Remarks on result:
other: Source, key, 68585-34-2, 1972
Dose descriptor:
NOAEL
Remarks:
developmental
Effect level:
>= 300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: No relevant treatment-related effects observed.
Remarks on result:
other: Source, key, 68585-34-2, 1972
Dose descriptor:
NOAEL
Remarks:
developmental
Effect level:
>= 165 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: No relevant treatment-related effects observed.
Remarks on result:
other: Source, key, 68585-34-2, 1972
Abnormalities:
no effects observed
Developmental effects observed:
no
Conclusions:
The NOAEL for the test substance was 300 mg/kg bw/day on the basis that the test substance did not produce a significant increase in the number of abnormal fetuses at any dose level. There were no significant test substance-related differences in the numbers of corpora lutea, implantation, resorptions or dead fetuses. Under the conditions of this study, the test substance, at the levels tested, was neither embryotoxic nor teratogenic.
Executive summary:

The developmental toxicity of the target substance is estimated based on an adequate and reliable prenatal developmental toxicity key study of a structural analogue source substance. The study used rabbits as test animals and revealed that the test substance did not produce a significant increase in the number of abnormal fetuses at any dose level. There were no significant test substance-related differences in the numbers of corpora lutea, implantation, resorptions or dead fetuses. Hence, under the conditions of the study, the test substance was neither embryotoxic nor teratogenic and the developmental NOAEL was determined to be > 300 mg/kg bw/day in rabbits. The results of the key study are further supported by additional (supporting) studies on varous analogue source substances. No relevant hazard with regard to developmental toxicity is therefore identified for the target substance as well. As explained in the category justification, the differences in molecular structure between the target and the source substances are unlikely to lead to differences with respect to developmental toxicity or teratogenicity.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subacute
Species:
rabbit
Quality of whole database:
The whole data base is conclusive and of high quality.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Data on developmental toxicity are available for various members of the AES category The AES reported within the category show similar structural, physico-chemical, environmental and toxicological properties. The approach of grouping different AES for the evaluation of their effects on human health and the environment was also made by the Danish EPA (2001) and HERA (2003), supporting the read-across approach between structurally related AES.

The purpose of the key study (BASF, 1994f) was to assess the effects of orally administered AES (C12-14) Na (CAS 68891-38-3) on embryonic and fetal development in pregnant CD rats. The study followed OECD Guideline 414 and complied with the OECD principles of GLP. In this study, AES (C12-14) Na (CAS 68891-38-3) was administered orally by gavage at dose levels of 0, 100, 300, and 1000 mg/kg bw/day once daily on Day 6-15 of gestation. In summary, the results of the study showed that repeated oral administration of AES (C12-14; 2EO) Na (CAS 68891-38-3) to pregnant rats did not cause symptoms of cumulative toxicity up to a dose level of 1000 mg/kg bw/day. There were no treatment-related fetal abnormalities at necropsy and no treatment-related effects in the reproduction data. Thus, based on the available information, the NOAEL for teratogenicity and developmental toxicity is assessed to be greater than 1000 mg/kg bw/day.

 

Pregnant albino rabbits, artificially inseminated, were administered AES (C10-16; 3 EO) Na (CAS 68585-34-2) by gavage at levels of 50, 100, or 300 mg/kg bw/day on Days 2-16 of gestation (P&G, 1972).No maternal deaths were attributable to the test substance and there were no significant differences in the number of corpora lutea, resorptions, implantations or live fetuses. The number of dead fetuses, 19 in the high level treatment group (300 mg/kg bw/day) and 17 in the control group, were higher than the remaining groups. All but four of these deaths occurred in just three litters. This lack of dose response indicated that something other than the test substance caused the deaths of these fetuses. In addition, no statistical differences were seen in the fetal weights of survivors or with regard to the number of fetuses with skeletal defects. The examination of fetuses from this study for soft-tissue abnormalities revealed only one instance of significant difference occurring in one litter of the low level treatment group. The lack of a dose response relationship between the levels of the test substance and the incidences of stressed bladder (five) indicate an isolated incidence of spontaneous malformations which have been seen in previous teratology studies. The NOAEL for the test substance was 300 mg/kg bw/day on the basis that the test substance did not produce any significant increases in the number of abnormal fetuses at any dose level. There were no significant test substance-related differences in the numbers of corpora lutea, implantation, resorptions or dead fetuses. Under the conditions of this study, the test substance, at the levels tested, was neither embryotoxic nor teratogenic.

 

References:

Danish EPA - Environmental and Health Assessment of Substances in Household Detergents and Cosmetic Detergent Products (2001). Environmental Project No. 615, pp. 24-28

HERA (2003). Human & Environmental Risk Assessment on ingredients of European household cleaning products Alcohol Ethoxysulphates, Human Health Risk Assessment Draft, 2003. http: //www. heraproject. com.

Justification for classification or non-classification

The available data on toxicity to reproduction do not meet the criteria for classification according to Regulation (EC) No. 1272/2008, and are therefore conclusive but not sufficient for classification.

Additional information