(E)-3-methyl-5-cyclopentadecen-1-one

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

concentration-driven

Effects on fertility

Description of key information

One-generation reproduction toxicity study: NOAEL (fertility) = 1000 mg/kg bw/d (OECD 415, GLP, K, rel. 1)

Link to relevant study records

Reference

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2001-12-11 to 2002-05-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study performed according to OECD test guideline No. 415 and in compliance with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

Inspected on 28 February 2000. Signed on 26 April 2000

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

The rat was selected as it is a readily available rodent species historically used in safety evaluation studies and acceptable to appropriate regulatory authorities

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS: Crl: CD (SD) IGS BR strain
- Source: Charles River (UK) Limited, Manston Road, Margate, Kent- UK
- Age at study initiation: not reported
- Weight at study initiation: Males: 184-265g; Females: 209-265g
- Fasting period before study: none
- Housing: Upon arrival, the animals were housed in groups of four in polypropylene cages with stainless steel grid floors and tops, suspended over paper-lined polypropylene trays. During the mating period animals were transferred to a similar type cage on a one male to one female per cage basis. Following evidence of successful mating, the males were returned to their original cages and transferred to a separate animal room of comparable conditions. The females were housed, individually, in polypropylene cages with solid floors and stainless steel tops. Mated females
were given softwood chips, as bedding throughout gestation and lactation.
- Diet (e.g. ad libitum): ad libitum (pelleted Rat and Mouse VRF1C Diet (Charles River UK Limited, Margate, Kent))
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 12 days (during which health status was assessed)


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 55 ± 15
(On isolated occasions the room temperature and/or humidity fell outside the protocol target limits but this was considered not to have affected the purpose or integrity of the study)
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 2001-12-11 To: 2002-05-15

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Test material was prepared at the appropriate concentration as a solution in 1.0% carboxymethyl cellulose (vehicle). For each dose level a separate aliquot of test material was weighed into the appropriate container. The vehicle was added and mixed using a Silverson homogeniser to ensure a homogeneous suspension was formed.

VEHICLE
- Justification for use and choice of vehicle (if other than water): no data
- Concentration in vehicle: 0, 5, 25 and 100 mg/L
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. (if required): no data
- Purity: no data

Details on mating procedure:

- M/F ratio per cage: 1 (1 female + 1 male per cage)
- Length of cohabitation: until evidence of successful mating (up to three weeks)
- Proof of pregnancy: The presence of sperm within the vaginal smear and/or vaginal plug in situ was taken as positive evidence of mating
- After successful mating each pregnant female was caged (how): Following evidence of successful mating, mated females were separated from male and housed individually during the period of gestation and lactation. The males were returned to their original cages and transferred to a separate animal room of comparable conditions.
- Any other deviations from standard protocol: none

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of ST 16 C 01 in the test material formulations was determined by gas chromatography (GC) using an external standard technique. Analysis of homogeneity, stability & concentrations in formulations gave good results, as shown hereafter:
HOMOGENEITY: The test material formulations were mixed thoroughly and samples were taken from the top, middle and bottom of the container, shaking in between sampling. Sampling was performed in triplicate. Measured concentrations were: 96.6-100.7%, 103.6-107.6% and 103.0-108.0% of nominal concentrations for 5, 25, 100 mg/ml nominal concentrations respectively.
STABILITY: The test material formulations were sampled and analysed initially and then after storage at approximately +4°C in the dark over the dosing period. Measured concentrations were 66-119%, 82-103%, 86-115 % respectively of nominal concentrations of 5, 25 and 100 mg/ml.
TEST MATERIAL FORMULATIONS: the test material formulations were sampled and analysed within weeks (Day 1, 8, 15, 22, 50, 78 & 107, 113, 115, 119) of preparation. Ranges of concentrations expressed in % of nominal concentration were 66-119%, 82-103% and 86-115% for the nominal concentrations of 5, 25 & 100 mg/ml respectively. Corresponding mean concentrations were 92%, 94 and 99 % for 5, 25 & 100 mg/ml nominal concentrations.

Duration of treatment / exposure:

(P) Males: at least 10 weeks before mating, then like females
(P) Females: at least 2 weeks prior to pairing, then throughout mating, gestation and lactation.

Frequency of treatment:

Daily, 7 days per week

Details on study schedule:

Male animals were dosed for at least ten weeks and female animals were dosed for at least two weeks, at their appointed dose levels, prior to pairing.
Parental males and females were paired within their respective dose groups for up to twenty one days.
Following evidence of mating, the animals were separated and males returned to their holding cages.
The pregnant females were allowed to deliver their offspring. The offspring were observed for growth and development during lactation.
At weaning on Day 21 (or as near to this date as possible) post partum the surviving offspring were killed and examined macroscopically post mortem.
The surviving adult Parental animals were killed and examined macroscopically post mortem.

Dose / conc.:

50 mg/kg bw/day (actual dose received)

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

28 (P) males caged with 28 (P) females per dose group for a total of 112 males and 112 females.
(see "Table 7.8.1/1: Animal assignment & doses" in "Any other information on material and methods including tables")

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The concentrations tested in this study were determined from a previous 28-d toxicity study (OECD 407 Guideline) performed with the same test material on the same rat strain and for which a NOEL of 250 mg/kg bw was determined.
- Rationale for animal assignment (if not random): random

Positive control:

Not used

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily during the normal week and once daily the during week-end and public holidays

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily, immediately before, immediately after and one hour after dosing for clinical signs of toxicity

BODY WEIGHT: Yes
- Time schedule for examinations: weekly during maturation and mating period. Following mating the parental males were weighed weekly until termination. Parental generation females showing evidence of mating were weighed on days 1, 4, 7, 14 and 21 post coitum. Parental generation females with a live litter were weighed on Days 1, 4, 7, 14 and 21 post partum.

FOOD CONSUMPTION:
During the maturation periods food consumption was recorded for each cage of parental generation adults. For parental generation females showing evidence of mating, food consumption was recorded for the periods covering days 1 to 7, 7 to 14 and 14 to 21 post coitum. For parental generation females with live litters, food consumption was recorded for the periods covering Days 1 to 7 and 7 to 14 postpartum.

Oestrous cyclicity (parental animals):

Each female was checked for the presence of copulation plug in vagina. A vaginal smear was prepared for each female and the stage of the oestrous cycle or the presence of sperm was recorded.

Sperm parameters (parental animals):

Not Applicable/Not required in OECD 415 Guideline

(P) males: weight of: epididymides, prostate, seminal vesicles (with coagulating gland), testes were determined.

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in offspring: Number and sex of live and dead pups, bodyweight/weight gain, clinical signs, physical or behavioural abnormalities, physical development (detachment of pinna, tooth eruption, eye opening), reflexological assessment (surface righting reflex, mid-air righting reflex, startle reflex and pupil reflex). For the time of observations, see Table 8.5.1/2 in "Any other information on material and methods including tables"


GROSS EXAMINATION OF DEAD PUPS:
yes, all offspring that died, or were killed in extremis during the lactation period were examined macroscopically and externally.

Postmortem examinations (parental animals):

All adult animals, killed in extremis or found dead during the course of the study, were examined macroscopically for internal and external abnormalities. All significant abnormalities were retained in fixative for possible further study.

SACRIFICE
- All animals: All surviving animals, following successful weaning of offspring, including non fertile animals.

GROSS NECROPSY
All animals were examined macroscopically for both internal and external abnormalities. Selected organs and tissues were retained in fixative.

HISTOPATHOLOGY / ORGAN WEIGHTS
Organ weights were determined for: Epididymides, Liver, Ovaries, Pituitary gland, Prostate, Seminal vesicles (with coagulating gland), Testes, Uterus with cervix.
Histopathology was performed on: Cervix, Coagulation gland, Epididymides, Liver, Pituitary gland, Prostate, Seminal vesicles, Significant abnormalities, Testes, Uterus, Vagina, Ovaries. (Initially the tissues from high dose and control animals were processed and embedded in paraffin wax BP (mp 56°C). Sections of the tissues were taken at 5 µm thickness, mounted on glass slides and stained with haematoxylin and eosin. In addition, any significant abnormalities from all dose groups were similarly processed.
Examination of the liver was extended to the intermediate and low dose groups.
The sections of reproductive and target organs from control and high dose animals were examined microscopically by a pathologist. Examination included an identification and characterisation of the individual cell stages of the seminiferous epithelium from sections of the testis.

Postmortem examinations (offspring):

All offspring that died, or were killed in extremis during the lactation period were examined macroscopically for externally.

SACRIFICE
- The F1 offspring were sacrificed at 21 days of age.
- These animals were subjected to postmortem macroscopic examinations for internal and external abnormalities.

Statistics:

The following parameters were analysed statistically, where appropriate using the test methods outlined as follows:
Adult male and female bodyweight during the maturation, gestation and lactation periods, adult male food consumption, female food consumption during maturation, gestation and lactation, litter size, litter weight, individual offspring bodyweight, offspring landmarks of physical development.
Values were analysed to establish homogeneity of group variances using Bartlett's test followed by oneway analysis of variance. If the variances were unequal subsequent comparisons between control and treated groups were performed using a standardised 'T' test. If variances were equal subsequent comparisons between control and treated groups were performed using Dunnett's Multiple Comparison Method.
Adult precoital intervals, female gestation lengths, offspring reflexological responses and litter sex ratios: Individual values were compared using the Kruskal-Wallis non-parametric rank sum test. Where significant differences were seen, multiple comparisons of control values .against treated group values was performed using Dunn's multiple comparison test.
Histopathology: Chi-squared analysis for differences in the incidence of lesions occurring with an overall frequency of 1 or greater. Kruskal-Wallis one-way non-parametric analysis of variance for the comparison of severity grades for the more frequently observed conditions.

Reproductive indices:

For each group the following were calculated.
Pre-coital interval: calculated as the time elapsing between initial pairing and the observation of positive mating
Mating Index % = number of animals mated/number of animals paired * 100
Pregnancy index % = Number of pregnant females / Number of animals mated * 100
Gestation length: calculated as the number of days of gestation including the days for observation of mating at the start of parturition. When the start of parturition occurred overnight, the total was adjusted by substracting half a day.
Parturition index % = Number of females delivering live pups / Number of pregnant females * 100

Offspring viability indices:

The following indices were calculated for each group from individual data.
Live birth index = Number of pups alive on Day 1 / Number of pups born * 100
Viability Index 1 (%) = Number of pups alive on Day 4/Number of pups alive on Day 1 * 100
Viability Index 2 (%) = Number of pups alive on Day 7/Number of pups alive on Day 4 * 100
Viability Index 3 (%) = Number of pups alive on Day 14/Number of pups alive on Day 7 * 100
Viability Index 4 (%) = Number of pups alive on Day 21/Number of pups alive on Day 14 * 100
Viability Index 5 (%) = Number of viable pups at weaning/Number of pups alive on Day 1 * 100

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

At 1000 mg/kg bw there was evidence of increased salivation, immediately post dosing for males and females. In addition, a significant number of males and females showed evidence of increased salivation prior to dosing. All other clinical findings were those commonly observed on this type of study. The frequency of findings was higher for males because of the longer dosing period. The observation of increased salivation is commonly associated with the oral administration of an unpleasant tasting test material and may not necessarily be indicative of systemic toxicity.
At 250 mg/kg bw there was evidence of increased salivation, particularly post dosing, for males and females. The incidence and/frequency were lower than for the highest dose level. There was a significant number of males with evidence of increased salivation pre dosing; although the incidence was lower than was observed at the highest dose level.
At 50 mg/kg bw there were isolated incidents of increased salivation although the incidence and frequency were low enough to be of no toxicological significance.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

At 1000 mg kg bw one male (number 94) was found dead and one male (number 99) was killed in extremis during the maturation phase of the study. One female (number 212) was killed in extremis during gestation. Male (number 94) had no significant clinical history prior to death. Male (number 99) and female (number 212) both showed evidence of ill-health, particularly respiratory distress, for 1-2 days prior to their demise. Post mortem studies showed macroscopic changes within the thoracic cavity affecting the lungs in particular. Histopathology showed evidence of pleuritis and/or abscess formation for all three animals. These macroscopic and microscopic changes are consistent with dosing trauma and not associated with systemic toxicity.
At 250 mg/kg bw two females (numbers 177 and 195) were found dead during lactation. There was no significant clinical history for either animal prior to death. There were no macroscopic or microscopic post mortem findings that could give a clear indication as to the likely cause of death for either animal.
At 50 mg/kg bw one male (number 30) was found dead during the maturation phase of the study. There was no clinical history prior to death. There were no macroscopic or microscopic post mortem findings that could give a clear indication as to the likely cause of death for this animal.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Maturation: there were no significant treatment-related effects upon male or female bodyweight gain and food consumption during the course of the maturation phase of the study. At 1000 mg/kg bw occasional week to week differences in male bodyweight gain compared to controls did not result in overall significant differences in group mean bodyweight.
Gestation & Lactation: there were no significant treatment-related effects upon female bodyweight gain and food consumption during gestation and lactation

Food consumption and compound intake (if feeding study):

no effects observed

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:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

At 1000 mg/kg bw there was a statistically significant increase in the incidence of hepatocyte enlargement for both male and female rats (p<0.001) when compared to control values. There were no significant microscopic changes observed amongst the reproductive organs of either sex.
At 250 mg/kg bw there was a statistically significant increase in the incidence of hepatocyte enlargement for both males and females (p<0.01 and pAt 50 mg/kg bw there was a statistically significant increase in the incidence of hepatocyte enlargement for females only (p<0.05) when compared to control values.

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

Fertility and Mating Performance: there were no significant treatment-related effects upon mating performance or fertility. The intergroup distribution of precoital intervals showed no significant treatment-related trends.
Gestation and Parturition: There were no significant treatment-related effects upon females or their offspring during pregnancy and parturition.

Key result

Dose descriptor:

NOAEL

Remarks:

fertility

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed

Key result

Critical effects observed:

no

Clinical signs:

no effects observed

Description (incidence and severity):

There were no significant treatment-related effects upon offspring clinical condition throughout lactation as indicated by comparable incidence and severity of clinical findings observed.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Description (incidence and severity):

There were no significant treatment-related differences in live birth index. There were no significant treatment-related differences in the live litter size and viability during lactation.
At 1000 mg/kg bw there was a reduction in offspring viability and therefore group mean litter size between days 7 and 14 of lactation. This was a result of significant offspring mortality amongst a small number of females. The isolated nature of these findings clearly shows that this is unrelated to dosing, particularly as offspring viability prior to this event was comparable to control values.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

There were no significant treatment-related effects upon individual offspring weight gain and physical development during lactation.
At 1000 mg/kg bw there was a reduction in group mean total litter weight on Day 14 of lactation. The difference was statistically significant (p<0.05) when compared to control values. The difference was most likely to be a consequence of the slight reduction in group mean live litter size. It was not a consequence of reduced offspring weight as group mean individual offspring values were comparable to controls.
At 250 mg/kg bw there was a slight reduction in group mean individual offspring bodyweight at day 7 of lactation only, the difference was statistically significant( p<0.05) when compared to control values. In addition, there was a delay in the group mean time of onset of pinna unfolding. The isolated nature of these differences and the lack of a dosage response clearly shows that these findings are not treatment-related.
At 50 mg/kg bw, there was a slight reduction in group mean individual offspring bodyweight at day 7 of lactation only. The difference was statistically significant (p<0.05) when compared to control values. There was a delay in the onset of eye opening. The difference was statistically significant (p<0.05) when compared to control values. The isolated nature of these differences and the lack of a dosage response clearly shows that these findings are not treatment-related.

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

Sexual maturation:

not examined

Anogenital distance (AGD):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no significant treatment-related differences in the type and incidence of macroscopic post mortem findings for offspring that died during the course of lactation or for offspring at terminal necropsy.

Histopathological findings:

not examined

Other effects:

no effects observed

Description (incidence and severity):

REFLEXOLOGICAL RESPONSE:
There were no significant treatment-related effects upon offspring reflexological responses.

SEX RATIO:
There were no significant treatment-related effects upon offspring sex ratios at Days 1 or Day 21 of lactation.

Key result

Dose descriptor:

NOAEL

Remarks:

growth, development and viability

Generation:

F1

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed

Key result

Critical effects observed:

no

Reproductive effects observed:

not specified

DISCUSSION: At 1000 mg/kg bw all mortalities showed evidence of dosing trauma as characterised by the macroscopic and microscopic changes to the thorax and lungs. This event is observed with the oral administration of test materials but is not indicative of systemic toxicity. During the in-life phase of the study the most notable finding at this dose level was the increased salivation; both pre and post dosing. This finding is a commonly observed reaction to the oral administration of an unpleasant tasting test material. The frequency and incidence of this finding shows a clear dose response. A slight difference in group mean bodyweights for males compared to control values was without statistical and toxicological significance. These findings show that the test material was well tolerated by adults of either sex during the course of the study. Post mortem evaluation did show a significant increase in liver weight for males. Histopathology showed an increased incidence of centrilobular hepatocyte enlargement. The effects upon the liver are commonly observed with the administration of xenobiotics and may therefore be considered an adaptive response rather than an adverse effect. There were no significant effects upon fertility or mating performance of adults.The live litter size at birth was comparable to control values. There was a reduction in offspring viability between lactation Days 7 and 14 and resulted in a slightly lower mean litter size between Days 14 and 21. The difference was not statistically significant and the reduction of offspring viability was the result of significant offspring mortality from a small number of litters. Offspring viability before and after this period was unaffected. It is therefore likely that the reduction in offspring viability was fortuitous and unrelated to treatment.

At 250 mg/kg bw there was no evidence to suggest that the small number of mortalities were a result of systemic toxicity. As was seen at the highest dose level, the most notable finding was the increased salivation, pre or post dosing. The incidence and frequency was lower than was seen at the highest dose level. There were no other significant effects on adults during the in-life phase of the study. Post mortem findings showed an elevation in liver weight for males only and a significant increase in hepatocyte enlargement for both sexes.These findings are consistent with those observed at the highest dose level. There was no effect upon reproductive performance and fertility of adults. The live birth index was comparable to controls and subsequent offspring growth and viability was unaffected by treatment. There were isolated differences in offspring growth or development compared to controls but these were without a true dosage response and without any consistent effect.

At 50 mg/kg bw there was one mortality which is unlikely to be a result of systemic toxicity. The low incidence of post dosing salivation could be fortuitous. There were no significant effects on adults during the in-life phase of the study. Post mortem findings were restricted to an increase in the incidence of centrilobular hepatocyte enlargement for females only. There were no significant effects upon the fertility and reproductive performance of adults. Offspring live litter size at birth, and subsequent growth and viability were comparable to control values. Isolated differences in offspring bodyweight and physical development which were statistically significant were considered not to be toxicologically significant because of the lack of dose response and the inconsistency of the findings.

Conclusions:

There were no toxicologically significant effects upon reproductive performance or fertility and no effects upon offspring viability, growth and development. The No Observed Adverse Effect Level (NOAEL) for fertility and offspring development is therefore 1000 mg/kg bw/d.

Executive summary:

In a one-generation reproduction toxicity study performed in accordance with OECD test guideline No. 415 and in compliance with GLP, the test material diluted in carboxymethylcellulose (CMC) was administered orally, by gavage, to groups of twenty-eight male and twenty-eight female rats throughout maturation, mating, gestation and lactation. The dose levels were 50, 250 and 1000 mg/kg bw/d with a similar sized control group receiving vehicle alone.

Following at least ten and two weeks of dosing respectively, male and female rats were paired within their dose groups to produce litters. At weaning of the offspring, all surviving animals were killed and examined macroscopically.

Parental animals were observed daily for clinical signs. Bodyweights and food consumption were recorded weekly during the maturation phase which was continued for males after the mating phase. Mated females were weighed and food consumption recorded on specific days post coitum and post partum.

The offspring were observed daily for clinical signs. The litter size and individual pup bodyweights were recorded on specific days post partum. During the lactation period, the offspring were observed for intra-litter onset and duration of landmarks or physical development. On specific days of lactation, reflexological assessment of offspring was performed.  

Post mortem macroscopic examinations were performed on all adults and offspring, including decedents. Reproductive and potential target organs were weighed from all parental animals and were then preserved in fixative. Histopathology was carried out on reproductive and target organs from control and high dose groups parental animals. Histopathology was extended to the low and intermediate dose groups for the liver (target organ) only.

 

At 1000 mg/kg bw/d in-life effects upon adults were limited to treatment-related increased salivation, pre and post dosing. There were no effects on fertility or reproductive performance.

Post mortem studies showed increased liver weights for males and an increased incidence of hepatocyte enlargement for both sexes. There were no effects upon reproductive organs.

At 250 mg/kg bw/d there was evidence of pre and post dosing salivation but at a lower incidence and frequency than at 1000 mg/kg. Post mortem studies showed an increased liver weight for males only and an increased incidence of hepatocyte enlargement for both sexes. There were no effects upon fertility or reproductive performance.

At 50 mg/kg bw/d there were isolated incidents of post dosing salivation but their relationship with test material administration are questionable when based on their frequency. There were no effects upon adult fertility and reproductive performance. Post mortem studies showed an increased incidence of centrilobular hepatocyte enlargement for females only.

 

There were no significant treatment-related effects upon live litter size at birth and throughout lactation. A reduction in offspring viability at 1000 mg/kg bw between days 7 and 14 of lactation was considered incidental as it was limited to a small number of females only. There were no significant treatment related differences in offspring growth and physical development during lactation. The statistically significant differences between control values and treated groups were incidental and unrelated to treatment.

 

The administration of the test material to male and female rats throughout one reproductive cycle at dose levels up to 1000 mg/kg bw/d resulted in adult toxicity that can be generalised as adaptive, non-specific responses to the administration of a xenobiotic.

There were no toxicologically significant effects upon reproductive performance or fertility and no effects upon offspring viability, growth and development.

The No Observed Adverse Effect Level (NOAEL) for fertility and offspring development is therefore 1000 mg/kg bw/d.

 

Under the test conditions, the test material is not classified according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

The study is GLP compliant and of high quality (Klimisch score = 1).

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 key study was identified (Safepharm, 2003, rel. 1). In this one-generation reproduction toxicity study performed in accordance with OECD test guideline No. 415 and in compliance with GLP, the test material diluted in carboxymethylcellulose (CMC) was administered orally, by gavage, to rats throughout maturation, mating, gestation and lactation. The dose levels were 50, 250 and 1000 mg/kg bw/d with a similar sized control group receiving vehicle alone. Following at least ten and two weeks of dosing respectively, male and female rats were paired within their dose groups to produce litters.

 

At 1000 mg/kg bw/d in-life effects upon adults were limited to treatment-related increased salivation, pre and post dosing. There were no effects on fertility or reproductive performance. Post mortem studies showed increased liver weights for males and an increased incidence of hepatocyte enlargement for both sexes. There were no effects upon reproductive organs.

At 250 mg/kg bw/d there was evidence of pre and post dosing salivation but at a lower incidence and frequency than at 1000 mg/kg. Post mortem studies showed an increased liver weight for males only and an increased incidence of hepatocyte enlargement for both sexes. There were no effects upon fertility or reproductive performance.

At 50 mg/kg bw/d there were isolated incidents of post dosing salivation but their relationship with test material administration are questionable when based on their frequency. There were no effects upon adult fertility and reproductive performance. Post mortem studies showed an increased incidence of centrilobular hepatocyte enlargement for females only.

 

There were no significant treatment-related effects upon live litter size at birth and throughout lactation. A reduction in offspring viability at 1000 mg/kg bw between days 7 and 14 of lactation was considered incidental as it was limited to a small number of females only. There were no significant treatment related differences in offspring growth and physical development during lactation. The statistically significant differences between control values and treated groups were incidental and unrelated to treatment.

 

The administration of the test material to male and female rats throughout one reproductive cycle at dose levels up to 1000 mg/kg bw/d resulted in adult toxicity that can be generalised as adaptive, non-specific responses to the administration of a xenobiotic.

There were no toxicologically significant effects upon reproductive performance or fertility and no effects upon offspring viability, growth and development.

The No Observed Adverse Effect Level (NOAEL) for fertility is therefore 1000 mg/kg bw/d. The NOAEL for growth, development and viability of offsprings is 1000 mg/kg bw/d.

Effects on developmental toxicity

Description of key information

This study is not required at REACh Annex VIII level.

In the One-generation reproduction toxicity study: NOAEL (growth, development and viability of offsprings) = 1000 mg/kg bw/d (OECD 415, GLP K, rel. 1)

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The study is GLP compliant and of high quality (Klimisch score = 1).

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

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No 1272/2008 (CLP).

Self-classification:

Based on the available information, no additional classification is proposed according to the CLP and to the GHS.