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Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

A study was conducted according to OECD guideline 421 in compliance with GLP in 2009. Ten Wistar rats per sex per dose were treated orally by gavage to 15, 40, and 100 mg/kg/day (incorporating a correction factor for 31.8% purity). No mortality was observed neither in the parental animals nor in the pups. No treatment-related effects were observed. The NOEL was determined to be 100 mg/kg/day.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
Jul 1995
GLP compliance:
yes (incl. certificate)
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: PA19100368
- Expiration date of batch: 31 Jan 2011
- Purity: 31.8%
- Physical appearance: pale amber liquid

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: The formulations were stable for at least 14 days
- Reactivity of the test substance with the solvent: Prepared formulations were within +/- 2% of the nominal concentration

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test substance was prepared as solution in distilled water
Species:
rat
Strain:
Wistar
Remarks:
Han:HsdRccHan:WIST
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: 12 weeks (parental male and females)
- Weight at study initiation: (P) Males: 305 - 358 g; Females: 189 - 219 g
- Housing: groups of five (initiation); one male: one female (mating); individually housed females (gestation and lactation)
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 2
- Humidity (%): 55 +/- 15
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 15 Sep 2009 To: 08 Nov 2009
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test material was administered daily using a stainless steel cannula attached to a disposable plastic syringe. Control animals were treated in an identical manner with 5 ml/kg/day of distilled water.
The volume administered to each animal was based on the most recent body weight and was adjusted at regular intervals.

Duration of treatment / exposure:
up to 55 consecutive days (including a two week maturation phase, pairing, gestation and early lactation for females).
Frequency of treatment:
daily (except for females during parturition where applicable)
Details on study schedule:
- Groups of ten male and ten female animals were treated daily. The first day of dosing was designated as Day 1 of the study
- F0 parental animals were mated on day 15 on a 1 male: 1 female basis
- Following evidence of mating (day 0 post coitum), males were returned to their cages and females were housed individually
- successful mating was assumed when the presence of sperm in the vaginal smear and/or vaginal plug in situ was observed
- pregnant females were allowed to give birth and maintain their offpsring until day 5 post partum
- males were killed and examined macroscopically on day 43
- females and surviving offspring were killed and examined macroscopically on day 5 post partum
Dose / conc.:
15 mg/kg bw/day
Remarks:
incorporating a correction factor for 31.8% purity
Dose / conc.:
40 mg/kg bw/day
Remarks:
incorporating a correction factor for 31.8% purity
Dose / conc.:
100 mg/kg bw/day
Remarks:
incorporating a correction factor for 31.8% purity
No. of animals per sex per dose:
10
Control animals:
yes, concurrent no treatment
Positive control:
no positive control used
Parental animals: Observations and examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: immediately before dosing, soon after dosing and one and five hours after dosing (working week); immediately before dosing, soon after dosing and one hour after dosing (weekends)

BODY WEIGHT: Yes
- Time schedule for examinations: on day 1 (prior to dosing) and then weekly until termination (males); weekly until mating was evident (females); on day 0, 7, 14 and 20 post coitum, and on day 1 and 4 post partum

FOOD CONSUMPTION:
- Food consumption for each animal determined: Yes
- Time schedule for examinations: weekly (during maturation period) until after mating phase (males); days 0-7, 7-14 and 14-20 post coitum (females) and during lactation period (days 1-4)

WATER CONSUMPTION: Yes
- Time schedule for examinations: daily by visual inspection
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, clinical condition from birth to day 5 post partum, individual offspring and litter weights on day 1 and 4 post partum, surface righting reflex

GROSS EXAMINATION OF DEAD PUPS:
yes (external and internal examinations)
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals on day 43
- Maternal animals: All surviving animals on day 5 post partum; females that failed to achieve pregnancy were killed after day 26 post coitum

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

HISTOPATHOLOGY
coagulation gland, epididymides, ovaries, mammary tissue, pituitary, prostate, seminal vesicles, testes, uterus/cervix, vagina
Postmortem examinations (offspring):
SACRIFICE
- all animals including those dying during the study

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations

HISTOPATHOLOGY:
coagulation gland, epididymides, ovaries, mammary tissue, pituitary, prostate, seminal vesicles, testes, uterus/cervix, vagina
Statistics:
- Data were assessed for dose response relationships by linear regression analysis, followed by one way analysis of variance (ANOVA) incorporating Levene’s test for homogeneity of variance. Dunnett's test was used for pairwise comparisons (variances homogenous). Non-parametric methods (Kruskal-Wallis ANOVA and Mann-Whitney "U" test) were used where Levene's test showed unequal variances and to analyze implantation loss, offspring sex ratio and ladmark developmental markers.
- p < 0.05 (significant = *)
- p > 0.05 (not significant)
- Chi-squared analysis (histopathology)
- Kruskal-Wallis one-way non-parametric analysis of variance (histopathology)
Reproductive indices:
Mating performance and fertility:
- pre-coital interval
- fertility indices (mating index, pregnancy index)
Gestation and parturition data:
- gestation length
- parturition index
Litter responses:
- implantation losses (pre-implantation loss, post-implantation loss)
Live birth and viability indices
- live birth index
- viability index
Sex ratio (%males)
Clinical signs:
no effects observed
Description (incidence and severity):
No toxicologically significant findings were observed throughout the study.
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
A slight but statistically significant increase in bodyweight gain (P<0.05) was evident in the 40 and 100 mg/kg/day males during Weeks 5 and 6. This increase was also seen in the 15 mg/kg/day males during Week 6 only. This finding is considered to be of no toxicological significance.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No adverse effect on dietary intake was detected for treated animals, in comparison to controls.
Food efficiency:
no effects observed
Description (incidence and severity):
No adverse effect on dietary intake was detected for treated animals, in comparison to controls.
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Daily visual inspection of the water bottles showed no adverse effect on water consumption.
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:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
No adverse effect on fertility was evident in the treated animals, in comparison to controls.
Two females (one in the 40 mg/kg/day and one in the 100 mg/kg/day) were not pregnant and displayed no corpora lutea and implantation sites. One female (40 mg/kg/day) had a total litter loss on pre-day 1 of lactation. Slight bilateral testicular atrophy was seen for one male which may have contributed to reduced reproductive performance and consequential non-pregnancy for one female.

There was no adverse effect on gestation lengths.
Dose descriptor:
NOEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed
Critical effects observed:
no
Clinical signs:
not examined
Description (incidence and severity):
No treatmentrelated clinical signs were detected.
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
At terminal kill, one female treated with 15 and 100 mg/kg/day produced a litter with one pup which showed a dark left testis. One 40 mg/kg/day female produced a litter with one pup which showed a dark right testis.
Histopathological findings:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
No obvious effect on litter size, sex ratio and offspring viability were detected from treated animals, in comparison to controls. No adverse effect on total litter weights, offspring bodyweight development or surface righting reflex was detected in treated animals, in comparison to controls.
Dose descriptor:
NOAEC
Generation:
F1
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed
Critical effects observed:
no
Reproductive effects observed:
not specified
Conclusions:
The oral administration of the substance to rats for a period of up to fifty-four consecutive days at dose levels of up to 100 mg/kg/day, did not result in any treatment-related effects. The ‘No Observed Effect Level’ (NOEL) for systemic toxicity was therefore considered to be 100 mg/kg/day (incorporating a correction factor for 31.8% purity). The ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 100 mg/kg/day (incorporating a correction factor for 31.8% purity).
Executive summary:

The study was performed to screen for potential adverse effects of the test material on reproduction including offspring development and provides an initial hazard assessment for effect on reproduction. The study complies with the recommendations of the OECD Guidelines for Testing of Chemicals No. 421 “Reproduction/Developmental Toxicity Screening Test” (adopted 27 July 1995).

The test material was administered by gavage to three groups, each of ten male and ten female Wistar Han™:HsdRccHan™:WIST strain rats, for up to fifty five consecutive days, (including a two week maturation phase, pairing, gestation and early lactation for females) at dose levels of 15, 40 and 100 mg/kg/day (incorporating a correction factor for 31.8% purity). A control group of ten males and ten females was dosed with vehicle alone (Distilled water). Clinical signs, bodyweight development, dietary intake and water consumption were monitored during the study.

Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 15 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex.

Adult males were terminated on Day 43, and all surviving females and offspring on Day 5 post partum. Any females that failed to achieve pregnancy were killed after Day 26 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed.

Results.

Adult Responses:

Mortality: There were no unscheduled deaths.

Clinical Observations: No toxicologically significant findings were observed throughout the study.

Bodyweight: No toxicologically significant effect on bodyweight development was detected for treated animals, in comparison to controls.

Food Consumption and Food Efficiency: No adverse effect on dietary intake was detected for treated animals, in comparison to controls.

Water Consumption: There was no adverse effect on water consumption for treated animals, in comparison to controls.

Reproductive Screening:

Mating, Gestation and Fertility: There were no intergroup differences in mating performance or gestation lengths. Pregnancy was achieved for ten control and 15 mg/kg/day females and eight 40 and 100 mg/kg/day females.

Litter Responses:

Offspring Litter Size, Sex Ratio and Viability: No adverse effects on litter size and viability were evident for treated animals, in comparison to controls. There was no obvious difference in sex ratio for litters from treated females, in comparison to controls.

Offspring Growth and Development: There was no adverse effect on offspring growth, bodyweight development or surface righting detected for treated animals, in comparison to controls.

Offspring Observations: The incidental findings detected in the control and treated groups were considered to be unrelated to treatment.

Pathology:

Necropsy: The incidental signs detected in the adults and offspring were considered to be of no toxicological importance.

Uterine Examination. From evaluation of the corpora lutea and implantation data, two females treated with 40 and 100 mg/kg/day showed no corpora lutea and implantation sites, therefore were never pregnant.

Organ Weights: No adverse effect on organ weight measurement was detected for treated animals, in comparison to controls.

Histopathology: There were no treatment-related changes in the reproductive organs examined.

 

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
Study duration:
subacute
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

A study was performed to screen for potential adverse effects of the test material on reproduction including offspring development and provides an initial hazard assessment for effect on reproduction. The study complies with the recommendations of the OECD Guidelines for Testing of Chemicals No. 421 “Reproduction/Developmental Toxicity Screening Test” (adopted 27 July 1995).

The test material was administered by gavage to three groups, each of ten male and ten female Wistar Han™:HsdRccHan™:WIST strain rats, for up to fifty five consecutive days, (including a two week maturation phase, pairing, gestation and early lactation for females) at dose levels of 15, 40 and 100 mg/kg/day (incorporating a correction factor for 31.8% purity). A control group of ten males and ten females was dosed with vehicle alone (Distilled water). Clinical signs, bodyweight development, dietary intake and water consumption were monitored during the study.

Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on Day 15 of the study, with females subsequently being allowed to litter and rear their offspring to Day 5 of lactation. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex.

Adult males were terminated on Day 43, and all surviving females and offspring on Day 5 post partum. Any females that failed to achieve pregnancy were killed after Day 26 post coitum. All animals were subjected to a gross necropsy examination and histopathological evaluation of reproductive tissues was performed.

RESULTS

Adult Responses:

Mortality: There were no unscheduled deaths.

Clinical Observations: No toxicologically significant findings were observed throughout the study.

Bodyweight: No toxicologically significant effect on bodyweight development was detected for treated animals, in comparison to controls.

Food Consumption and Food Efficiency: No adverse effect on dietary intake was detected for treated animals, in comparison to controls.

Water Consumption: There was no adverse effect on water consumption for treated animals, in comparison to controls.

Reproductive Screening:

Mating, Gestation and Fertility: There were no intergroup differences in mating performance or gestation lengths. Pregnancy was achieved for ten control and 15 mg/kg/day females and eight 40 and 100 mg/kg/day females.

LITTER RESULTS:

Offspring Litter Size, Sex Ratio and Viability: No adverse effects on litter size and viability were evident for treated animals, in comparison to controls. There was no obvious difference in sex ratio for litters from treated females, in comparison to controls.

Offspring Growth and Development: There was no adverse effect on offspring growth, bodyweight development or surface righting detected for treated animals, in comparison to controls.

Offspring Observations: The incidental findings detected in the control and treated groups were considered to be unrelated to treatment.

PATHOLOGY:

Necropsy: The incidental signs detected in the adults and offspring were considered to be of no toxicological importance.

Uterine Examination. From evaluation of the corpora lutea and implantation data, two females treated with 40 and 100 mg/kg/day showed no corpora lutea and implantation sites, therefore were never pregnant.

Organ Weights: No adverse effect on organ weight measurement was detected for treated animals, in comparison to controls.

Histopathology: There were no treatment-related changes in the reproductive organs examined.

 

The analogue Amides, C12-C18 (even numbered), N-[3-(dimethylamino) propyl], N´-oxides which shares the same functional groups with the substance Amides, C12-C14 (even numbered), N-[3-(dimethylamino) propyl], N´-oxides, also has comparable values for the relevant molecular properties.

Based on company experimental data (reported under the endpoint record Toxicity to reproduction.001) on the analogue, the read-across approach is applied and the NOEL for reproductive and systemic toxicity for the substance Amides, C12-14 (even numbered), N-[3-(dimethylamino)propyl], N'-oxides is considered to be 100 mg/kg/day.

 

Effects on developmental toxicity

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 study available

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for reproductive toxicity under Regulation (EC) No. 1272/2008.