Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
no study available
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

Polysulfides di-tert-dodecyl did not result in toxicity to the embryo or fetus or show developmental toxicity after gavage dosing (GD 6-15) to rats at doses as high as 1000 mg/kg bw/day in the developmental toxicity test (OECD TG 414). The NOAEL for both maternal and fetal toxicity was higher than 1000 mg/kg bw/day.

In an OECD TG 414 study, three groups of 25 mated female rats received polysulfides, di-tert-dodecyl by oral gavage at the dose levels of 50, 250 or 1000 mg/kg/day, each day from day 6 to day 15 post-coitum inclusive (Richard, 1997). Simultaneously, a group of 25 mated females was given the vehicle alone (0.5% carboxymethylcellulose) under the same conditions and acted as a control group. Clinical signs including (including evidence of abortion/resorption) and mortality were checked daily. Food consumption and body weight were recorded at designated intervals during pregnancy. On day 20 post-coitum, females were killed. The gravid uterus was weighed and fetuses were removed by hysterectomy. Females were examined macroscopically. Litter parameters were recorded: number of corpora lutea, implantation sites, resorptions, dead and live fetuses. The live fetuses were weighed, sexed, submitted to an external examination and then to soft tissue or skeletal examinations. No clinical signs and no unscheduled deaths were observed in any group. No females aborted or presented total resorption in any group. The food consumption and body weight gain of the pregnant females from all treated groups were similar to those of controls. No treatment-related macroscopic findings were observed, in any group. The post-implantation loss was similar in the 0, 50 and 250 mg/kg/day groups. In the 1000 mg/kg/day group, a slightly increased post-implantation loss (represented mainly by late resorptions, observed in one female) was observed: it could not be demonstrated that this single event was treatment-related. No treatment-related effects were observed on the number of live fetuses per animal, the fetal body weight or the sex-ratio. No treatment-related external, soft tissue and skeletal anomalies or malformations were observed in any group. The No Effect Level was higher than 1000 mg/kg/day in terms of maternotoxicity, embryofetotoxicity and teratogenic effects.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River France (76410 Saint-Aubin-lès-Elbeuf, France)
- Age at study initiation: 9 weeks old
- Weight at study initiation: mean body weight of 272 g (range: 235 g to 306 g)
- Fasting period before study: no
- Housing: individually in polycarbonate cages
- Diet (ad libitum): A04 C pelleted diet (UAR, France)
- Water (ad libitum): filtered tap wtaer
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 50 ± 20
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was prepared as a dispersion in the vehicle, in order to achieve the concentrations of 10, 50 and 200 mg/ml. The test substance was carefuliy mixed with the required quantity of vehicle using an ultraturrax mixer, for approximately two minutes and then homogenized using a magnetic stirrer.
The test substance preparations were made for up to seven days of treatment, on the basis of available stability data (9 days at +4 °C) and were stored at +4 °C, away from light, pending utilization.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration of samples of each dispersion (including the control) prepared for use on the first day of treatment of the first mated females and on the last day of treatment of the last mated females was determined by HPLC with UV detection.
Details on mating procedure:
Females were mated at the breeder's facilities.
The day of the positive mating (assessed by observation of vaginal plug) was designated as day 0 post-coitum (p.c.).
Duration of treatment / exposure:
Gestation day 6 to 15
Frequency of treatment:
daily
Duration of test:
up to gestation day 20
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Details on study design:
Dose selection rationale: based of the results of a four-week toxicity study (C.I.T./study No 12602 TSR): . No toxic effects were noted after four weeks, at dose-levels up to 1000 mg/kg/day.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day for clinical signs and at least twice a day for mortality

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: on days 2, 6, 7, 8, 9, 10, 12, 14, 16 and 20 post-coitum.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
on days 2-6, 6-9, 9-12, 12-16 and 16-20 post-coitum.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined:
After hysterectomy, the females were subjected to a macroscopic examination of the principal thoracic and abdominal organs.
Ovaries and uterine content:
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
- Other: number and distribution of dead and live fetuses
In apparently non-pregnant females, the presence of implantation sites was checked using the Salewski staining technique
Fetal examinations:
- External examinations: Yes: [all per litter ]
- Soft tissue examinations: Yes: [half per litter ]
- Skeletal examinations: Yes: [half per litter ]
- Head examinations: Yes: [all per litter ]
Statistics:
Mean values were compared by one-way analysis of variance and Dunnett's test, values being considered as normally distributed, variances being considered as homogenous. Percentage values were compared by Fischer's exact probability test.
Indices:
Pre-implantation loss; post-implantation loss
Clinical signs:
no effects observed
Description (incidence and severity):
No relevant clinical signs were observed in any female from the control, 50, 250 or 1000 mg/kg/day groups.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
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:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Dilated renal pelvis was observed in 1/25 females of the 50 mg/kg/day group (unilateral) and in 2/25 females of the 1000 mg/kg/day group (bilateral). This finding which was observed with a low, and not dose-related incidence and which could be observed spontaneously in female rats of this age and strain, was not attributed to the test substance administration.
The other macroscopic findings encountered did not show any indication of treatment or dose-relationship.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not specified
Number of abortions:
no effects observed
Pre- and post-implantation loss:
effects observed, non-treatment-related
Description (incidence and severity):
The mean number of corpora lutea per animal was similar in the control and treated groups (17.0, 18.2 or 17.7 at 50, 250 or 1000 mg/kg/day vs. 17.8 in controls).
The mean number of implantation sites per animal was similar in the control and treated groups (13.6, 15.1 or 12.8 at 50, 250 or 1000 mg/kg/day vs. 12.3 in controls).
The normal variations observed for the preimplantation loss were considered to be fortuitous since treatment started after implantation.
Accordingly, the post implantation loss was low and similar in the control, 50 or 250 mg/kg/day groups (3.3% or 5.0% of implantation sites at 50 or 250 mg/kg/day vs. 4.5% in controls) and slightly higher at 1000 mg/kg/day (10.7% of implantation sites at 1000 mg/kg/day vs. 4.5% in controls; difference statistically significant: p < 0.05).
This value was slightly outside the range of the historical control data: average = 3.9%, mini = 0.8%, maxi = 7.7% (period: 1995-1996; 202 females evaluated).
Since this difference from controls was mainly attributable to a single individual, and since no other signs of embryo or fetotoxicity were observed, it could not be demonstrated that this single event was treatment-related.
Total litter losses by resorption:
not examined
Early or late resorptions:
effects observed, non-treatment-related
Description (incidence and severity):
The mean number of resorptions (early and late) per animal was low and similar in the control and 50 or 250 mg/kg/day groups (0.4 or 0.8 at 50 or 250 mg/kg/day vs. 0.6 in controls, corresponding to a rate of 3.3 or 5.0% of implantation sites, respectively vs. 4.5% in controls). At 1000 mg/kg/day, the mean number of resorptions (early and late) per animal was slightly higher (1.3 vs. 0.6 in controls corresponding to a rate of 10.2% of implantation sites vs. 4.5% in controls; difference statistically significant for the rate: p < 0.05); this difference from controls was due to higher rate of late resorption (6.8% of implantation sites vs. 0.9% in controls; difference statistically significant: p < 0.01) attributed to only one female (13 late resorptions out of 16 implants, female Q24440).
Dead fetuses:
effects observed, non-treatment-related
Description (incidence and severity):
No dead fetuses were noted in the control, 50 or 250 mg/kg/day groups.
In the high dose-level group, only one fetus was found dead in the female which presented a high rate of resorptions.
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
no effects observed
Other effects:
not specified
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Fetal body weight changes:
no effects observed
Description (incidence and severity):
The mean fetal body weight was similar in the control and treated groups (4.00, 3.95 or 4.07 g at 50, 250 or 1000 mg/kg/day, respectively, vs. 3.91 g in controls).
Reduction in number of live offspring:
effects observed, non-treatment-related
Description (incidence and severity):
The mean number of live fetuses per animal was similar in the control and treated groups (13.2, 14.3, or 11.4 at 50, 250 or 1000 mg/kg/day, respectively, vs. 11.8 in controls). The rate of live fetuses was slightly lower in the 1000 mg/kg/day group, when compared to the control group, as a consequence of the higher post-implantation loss.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
The sex-ratio was similar in the control and treated groups, and close to the theoretical value of 50%.
External malformations:
no effects observed
Description (incidence and severity):
No external anomalies or malformations were observed in the fetuses of the 50, 250 or 1000 mg/kg/day groups. In the control group, two fetuses (from the same female) presented malformations.
Skeletal malformations:
no effects observed
Description (incidence and severity):
. Malformations
No skeletal malformations were noted in any fetus of the control, 50 or 250 mg/kg/day group. Fused and distorted lumbar was noted in 1/94 fetuses of the high dose-level group. Since this malformation was noted with a very low incidence and was not associated with any other skeletal anomaly or variation, it was considered to be of spontaneous occurrence.
. Anomalies
The total fetal incidence of skeletal anomalies was similar in all control and treated groups i.e. 36/137 (26.3%) at 50 mg/kg/day, 49/158 (31.0%) at 250 mg/kg/day or 24/94 (25.5%) at 1000 mg/kg/day v.s. 34/109 (31.2%) in controls. No relevant or dose-related differences from controls were noted in any recorded anomalies, and ail were commonly recorded findings in fetuses of this rat strain.
. Variations
The total fetal incidence of skeletal variations was similar in all control and treated groups i.e. 107/137 (78.1%) at 50 mg/kg/day, 120/158 (75.9%) at 250 mg/kg/day or 70/94 (74.5%) at 1000 mg/kg/day v.s. 91/109 (83.5%) in controls. No dose-relationship and no statistically significant difference from controls were noted for the principal variations observed (reduced or absence of ossification of sternebrae).
Visceral malformations:
no effects observed
Description (incidence and severity):
. Malformations
No fetal soft tissue malformations were noted in any fetus of any treated group. The two control fetuses malformed at external examination presented several internal soft tissue malformations.
. Anomalies
The litter and the fetal incidences of the few recorded anomalies were similar in control and treated groups, without dose-relationship and close to the higher limit or in the range of the historical control data. Consequently, they were not considered to be treatment-related, but to be of a spontaneous nature.
Other effects:
not specified
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Developmental effects observed:
not specified

Table 1 : pregnancy status

Dose-levels (mg/kg/day)

0

50

250

1000

Number of mated females

25

25

25

25

Not pregnant

7

5

4

9

Pregnant

18

20

21

16

. found dead

0

0

0

0

.aborted

0

0

0

0

Alive at terra

18

20

21

16

. total resorption

0

0

0

0

Completed pregnancy

18

20

21

16

Conclusions:
TPS 32 , administered daily by the oral route at 50, 250 or 1000 mg/kg/day to pregnant Sprague-Dawley female rats during organogenesis did not show any signs of toxicity in the pregnant female and did not produce any embryotoxicity, fetotoxicity or teratogenic effects. At 1000 mg/kg/day, only a slight increase in post-implantation loss was noted, as a contribution of a single female: it could not be demonstrated that this single event was treatment-related. The No Effect Level is defined as 1000 mg/kg/day in terms of maternotoxicity, embryofetotoxicity and teratogenic effects.
Executive summary:

In an OECD TG 414 study, three groups of 25 mated female rats received di-tert-dodecyl polysulfides (TPS 32), by oral gavage at the dose levels of 50, 250 or 1000 mg/kg/day, each day from day 6 to day 15 post-coitum inclusive. Simultaneously, a group of 25 mated females was given the vehicle alone (0.5% carboxymethylcellulose) under the same conditions and acted as a control group.Clinical signs including (including evidence of abortion/resorption) and mortality were checked daily. Food consumption and body weight were recorded at designated intervals during pregnancy. On day 20 post-coitum, females were killed. The gravid uterus was weighed and fetuses were removed by hysterectomy. Females were examined macroscopically. Litter parameters were recorded: number of corpora lutea, implantation sites, resorptions, dead and live fetuses. The live fetuses were weighed, sexed, submitted to an external examination and then to soft tissue or skeletal examinations.

No clinical signs and no unscheduled deaths were observed in any group. No females aborted or presented total resorption in any group. The food consumption and body weight gain of the pregnant females from all treated groups were similar to those of controls. No treatment-related macroscopic findings were observed, in any group.

The post-implantation loss was similar in the 0, 50 and 250 mg/kg/day groups. In the 1000 mg/kg/day group, a slightly increased post-implantation loss (represented mainly by late resorptions, observed in one female) was observed: it could not be demonstrated that this single event was treatment-related. No treatment-related effects were observed on the number of live fetuses per animal, the fetal body weight or the sex-ratio.No treatment-related external, soft tissue and skeletal anomalies or malformations were observed in any group. 

The No Effect Level is defined as 1000 mg/kg/day in terms of maternotoxicity, embryofetotoxicity and teratogenic effects.
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
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Toxicity to reproduction: other studies

Description of key information

In an OECD TG 407 study (Molinier, 1997), male and female Sprague-Dawley rats administered 0, 50, 250, or 1000 mg/kg bw/day of polysulfides, di-tert-dodecyl by daily gavage for 29 days. No effects on reproductive organ weights or histopathology were observed at 1000 mg/kg bw/day, the highest dose tested.

Justification for classification or non-classification

According to the available data and criteria of Regulation no. 1272/2008, no classification is warranted for reproductive and developmental toxicity.