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Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Two screening studies for reproduction and developmental effects were performed according to OECD 422 with two representatives of the analogue Amphoacetates C8 -C18 (a mono- and a diacetate form, respectively). This data is read across to Amphoacetates C8, the rationale is attached in Section 13.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
The rationale to read across the data is attached in Section 13.
Reason / purpose for cross-reference:
read-across source
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Animals of the 1000 mg/kg group and, on only a few occasions, in animals of the 300 mg/kg group:
Observed salivation was considered to be a physiological response rather than a sign of systemic toxicity considering its slight severity and the time of occurrence (i.e. after dosing).
Rales were noted in several males and females of the 1000 mg/kg group (and in a single male and female of the 100 mg/kg group). As these animals showed rales on only one or a few days, this finding was considered not to be toxicologically relevant (it was likely related to the dosing technique). Any other clinical signs noted incidentally occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study and showed no dose-related trend. At the incidence observed, these were considered to be unrelated to treatment.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One male and two females died in the highest dose group. The male was euthanized in extremis on day 17 of the treatment period (day 3 of mating period). Body weight gain was normal up to day 15. Necropsy showed macroscopic findings such as mucous contents in the trachea and gas distended parts of the gastrointestinal tract. Microscopic findings showed acute inflammation of the trachea. One female was found dead on day 16 of the treatment (day 1 of the post-coitum period). The weight gain during the whole period was low. Macroscopic findings were swollen lungs; microscopic findings were alveolar content and congestion of the lungs, marked bronchial mucosal erosion and erosion/ulceration of the trachea. One female was euthanized in extremis on day 40 of the treatment (day 1 of lactation period). Food consumption and weight gain was reduced since day 17-20 of the gestation period. Macroscopic findings were pale liver and greenish kidneys. Microscopic findings were marked ulceration in the forestomach and lymphogranulocytic inflammation. Mortality was considered to be associated to the dosing technique and not test-item related.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Mean body weights and body weight gain were considered not to be adversely affected by treatment.
A few findings at 1000 mg/kg bw/day were regarded as not toxicologically relevant as explained below.
Two males in the dose groups 1000 mg/kg bw/day showed considerably reduced body weight gain over the 4-week treatment period whereas the other males of this dose group grew normally. There were no associated signs of toxicity and mean body weights of 1000 mg/kg bw/day males remained close to control values (4% difference at the end of the treatment period, not statistically significant).
Findings of note in 1000 mg/kg females consisted of slightly lower mean body weight gain in the last week of the gestation period and reduced weight gain or slight weight loss in three females during the lactation period. Mean body weights of 1000 mg/kg bw/day females did not differ statistically significantly from those of controls (5% difference at the end of the post-coitum and lactation periods). Except for the female which was euthanized in extremis on Day 1 of the lactation period, the lower weight gain was not associated with signs of toxicity.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Females in the dosing groups of 1000 mg/kg showed reduced food consumption (before and after correction for body weight) in two periods: Periods between days 14-20 of the post-coitum period (about 10%, statistically significant) and during the lactation period (statistically significant between days 7-13; mean absolute food consumption in this interval was 20% lower than the control value).These findings were considered not to be toxicologically relevant as they were not associated with an adverse effect on body weight gain.
Food efficiency:
not specified
Description (incidence and severity):
Relative Food Consumption calculated against the body weight for scheduled intervals
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
There were no treatment-related changes in red blood cell parameters, white blood cell parameters or
number of platelets.
Males treated at 1000 mg/kg had slightly lower activated partial thromboplastin time (APTT) values than concurrent controls. As all values at 1000 mg/kg remained within the historical control range, this change was regarded as non-adverse.
Female rats showed no treatment-related changes in coagulation parameters.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Clinical chemistry parameters showed no differences between control and treated rats that were considered to be toxicologically significant.
Mean values for alanine aminotransferase (ALAT) and bile acids in males treated at 1000 mg/kg were higher compared to the control group means (40 and 77%, respectively). These differences were not statistically significant, mean values at dose group 1000 mg/kg remained in the historical control ranges, and there were no associated anatomic pathology alterations. As such, these clinical chemistry findings were regarded as non-adverse.
Isolated, statistically significant variations noted in clinical chemistry parameters (higher creatinine and sodium in males at 300 mg/kg) were considered to be unrelated to treatment due to the lack of a dose-related trend and/or small magnitude of the difference from controls.
Serum levels of T4 in males were not affected by treatment.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Hearing ability, pupillary reflex and static righting reflex were normal in all examined animals.
Grip strength was not affected by treatment. A statistically significantly higher forelimb grip strength noted in males at 300 mg/kg was judged to be unrelated to treatment due to the lack of a dose-related trend.
The variation in motor activity did not indicate a relation with treatment. All groups showed a similar habituation profile with a decreasing trend in activity over the duration of the test period. In the absence of a dose-related trend, the higher values for total movements and ambulations noted in 100 mg/kg females, particularly two of them, were regarded as unrelated to treatment.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
All of the recorded microscopic findings were within the range of background pathology encountered in rats of this age and strain. There was no test item-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.
Histopathological findings: neoplastic:
no effects observed
Other effects:
effects observed, non-treatment-related
Reproductive function: oestrous cycle:
effects observed, non-treatment-related
Description (incidence and severity):
Length and regularity of the estrous cycle were not affected by treatment. During the treatment (premating) period, all females had regular cycles of four days.
Extended di-estrus during pairing occurred in one female of the 100 mg/kg group which showed no evidence of mating. The irregular cycle noted in one female of the 1000 mg/kg group was not test item-related as it occurred prior to initiation of treatment.
Reproductive function: sperm measures:
effects observed, treatment-related
Description (incidence and severity):
One male at 300 mg/kg bw/day showed tubular atrophy in the testes and reduced luminal sperm with luminal cell debris in the epididymides which accounted for the lack of offspring. This male had no normal spermatogenic staging profile. For the other evaluated testes no indications for abnormal spermatogenesis were seen.
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
Seven couples had no offspring: 2/10 control couples (both females not pregnant); 2/10 couples at 100 mg/kg bw/day (one female without evidence of mating; one female not pregnant); 2/10 couples at 300 mg/kg bw/day (one female not pregnant; one female implantation sites only); 1/10 couples at 1000 mg/kg bw/day (one female not pregnant).
Key result
Dose descriptor:
NOAEL
Remarks:
Parental/ Reproduction
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No toxicity was observed up to the highest dose level tested.
Key result
Critical effects observed:
no
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Clinical signs were seen only in pups of the 1000 mg/kg bw/day group that did not survive until scheduled sacrifice. The findings in the pups of one litter (cold, no milk in the stomach) were related to the moribundity of the dam, which was not test item-related.
The nature of the findings in a few other pups (pale appearance, cold, no milk in the stomach) remained within the range seen normally in pups that die within a few days after birth. These findings were therefore regarded as unrelated to treatment.
Treated pups that survived until scheduled sacrifice showed no clinical signs and the incidental findings in pups of the control group (alopecia, pale appearance) remained within the range considered normal for pups of this age.
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
The number of live offspring in day 1 after littering compared with the total number of offspring born was considered not to be affected by treatment. At 1000 mg/kg bw/day, seven pups (from 4 litters) were found dead at first litter check. The pup mortality in one of the litters was considered to be related to the moribundity of the dam (euthanized in extremis at PND 1; her moribundity was not test item-related. The number of dead pups in the other 1000 mg/kg bw/day litters was within normal limits and judged to be unrelated to treatment.
The same was true for the incidental mortality at 100 mg/kg bw/day (one pup from 1 litter).The number of live offspring on PND 4 compared with the number of live offspring on PND 1 was considered not to be affected by treatment. At 1000 mg/kg bw/day, a total of 15 pups (out of 4 litters) died at PND 1-2 (versus none in the control group; the difference was statistically significant). These pups went missing, presumably cannibalized, died spontaneously, or were euthanized in extremis (10 pups of one dam that were euthanized at PND 1). The moribundity of the dam, resulting in poor health of her pups, was not test item-related. Pup mortality in the other 1000 mg/kg bw/day litters remained within the range considered normal for pups of this age and was therefore considered to be unrelated to treatment. For the same reason the incidental pup mortality at the lower dose levels was regarded as unrelated to treatment (one pup out of one litter at dose 100 mg/kg bw/day, two pups out of two litters at dose 300 mg/kg bw/day).
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weights of pups were not affected by treatment.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Serum T4 levels in male and female PND 14-16 pups were not affected by treatment. Assessment of T4 for PND 4 pups and TSH for PND 14-16 pups was considered not relevant because no treatment-related changes in T4 were noted in pups at PND 14-16.
Sexual maturation:
not examined
Anogenital distance (AGD):
no effects observed
Description (incidence and severity):
Anogenital distance (absolute and normalized for body weight) in male and female pups was not affected by treatment.
Nipple retention in male pups:
no effects observed
Description (incidence and severity):
Treatment up to 1000 mg/kg be/day had no effect on areola/nipple retention. For none of the examined male pups nipples were observed at PND 13.
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No macroscopic findings were noted among pups that were considered to be test item-related. Absence of milk in the stomach was noted in most of the pups found dead at first litter check. As absence of milk in the stomach is a normal finding in pups that die prematurely and the incidence among pups of surviving dams remained in the range considered normal for pups of this age, this macroscopic finding was regarded as unrelated to treatment. No other macroscopic findings were noted in pups that died prematurely. Pups that survived until scheduled sacrifice showed no abnormalities at macroscopic examination.
Histopathological findings:
not examined
Other effects:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Remarks:
Repro/ development
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 effects seen up to the highest dose level tested (1000 mg/kg bw/day)
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no

Analysis of dose preparations: The concentrations analyzed in the test item formulations were in agreement with target concentrations (i.e. mean accuracies between 93% and 100%). A small response noted at the retention time of the test item in the chromatograms of the control group formulation was considered to derive from carry-over in the analytical system. This had no significant effect on the results of the other study samples because of the minor magnitude (maximally 0.0092% relative to low dose group samples). The formulations of the low and the high dose group were homogeneous (i.e. coefficient of variation≤5.8%).

Conclusions:
In an oral OECD 422 screening study with substance analogue Amphoacetates C8-C18, the parental and reproductive NOAEL were derived to be 1000 mg/kg bw/day. No parental, reproduction and developmental toxicity was observed up to the highest dose level tested (1000 mg/kg bw/day). This result is read across to Amphoacetates C8.
Executive summary:

A combined oral repeated dose study with screening for reproductive and/ or developmental effects was performed according to OECD/EC guidelines and GLP principles with substance analogue Amphoacetates C8-C18. Miranol Ultra C32 was administered by daily oral gavage to male and female rats at dose levels of 100, 300 and 1000 mg/kg bw/day. Males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 29 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation (for 50-56 days). Treatment with Miranol Ultra C32 was associated with a few non-adverse changes at the highest dose group i.e. slight salivation in both sexes, lower food consumption in females in the last week of gestation and during lactation, and lower activated partial thromboplastin time in males. No treatment-related or toxicologically relevant changes were noted in the other parameters investigated in this study. Based on the absence of adverse effects up to 1000 mg/ kg bw/day, a parental No Observed Adverse Effect Level (NOAEL) for Miranol Ultra C32 of 1000 mg/kg bw/day was established. The NOAEL for reproduction was established to be 1000 mg/ kg bw/day. This result is read across to Amphoacetates C8.

Effect on fertility: 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 available studies are performed according to OECD/EC guidelines and GLP principles (Klimisch 1 studies).
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available

Effects on developmental toxicity

Description of key information

Two screening studies for reproduction and developmental effects were performed according to OECD 422 with two representatives of Amphoacetates C8-C18. One study was done with a mono-acetate form, the second study was done with a diacetate Amphoacetate C8-C18. The rationale to perform the test with both forms was to demonstrate that both substances are of low toxicity and to demonstrate that the toxicological hazard of both forms are covered in the registration. The data are read across to Amphoacetates C8 -C18.

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 available studies are performed according to OECD/EC guidelines and GLP principles (Klimisch 1 studies).
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

Two screening studies for reproduction and developmental effects were performed according to OECD 422 with two representatives of Amphoacetates C8-C18. In both studies, no reproduction toxicity was observed up to the highest dose level tested (1000 mg/kg bw/day). No treatment-related changes were noted in the reproductive parameters examined in both studies (i.e. mating and fertility indices, precoital time, number of implantation sites, estrous cycle, spermatogenic profiling, and histopathological examination of reproductive organs). For the study with the diacetate Amphoacetate C8-C18, the reproduction parameters were assessed for all groups, but the developmental effects could not be determined in the high dose group. In neither study developmental toxicity was observed up to the highest dose level tested (1000 mg/kg bw/day or 300 mg/kg bw/day). No treatment-related changes were noted in the developmental parameters investigated in the studies (i.e. gestation, viability and lactation indices, duration of gestation, parturition, sex ratio, maternal care and early postnatal pup development consisting of mortality, clinical signs, body weight, anogenital distance (PND 1), areola/nipple retention (PND 13 males), T4 thyroid hormone levels (PND 14-16) and macroscopy). Furthermore, no effects on weight, macroscopy or histopathology were seen in the sub-chronic study on male and female reproductive organs performed with substance analogue diacetate Amphoacetate C8-C18. Stage dependent qualitative evaluation of spermatogenesis in the testes was performed. The testes revealed normal progression of the spermatogenic cycle and the expected cell associations and proportions in the various stages of spermatogenesis were present. Taken together, there are no indications that the registered substance has an adverse effect on reproduction and development.

Justification for classification or non-classification

Based on the available data, the registered substance is not classified for reproduction toxicity and effects on development according to CLP Regulation (EC) No. 1272/2008.

Additional information