Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Multigeneration studies were available for read across substance Docusate sodium for assessment of reproductive and postnatal developmental function. In a first study, slight maternal/paternal toxicity were observed at 0.5 and 1% in the diet, however this was not confirmed in a second study. The test article was considered to have influenced the taste of the milk, for which adaptations were done in the second study. From both studies, it can be concluded that dosing Docusate sodium up to 1% in the diet did not lead to effects on fertility or postnatal development; this concentration corresponds with 750 mg/kg bw/day.
Based on the absence of reproductive findings in the repeated dose toxicity studies and the multigeneration studies, no further testing is needed.


Link to relevant study records

Referenceopen allclose all

Endpoint:
three-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
Read across argumentation is provided in Section 13.
Reason / purpose:
read-across source
Clinical signs:
no effects observed
Description (incidence and severity):
There were no treatment-related clinical observations for F0 males and females. Alopecia was noted with similar frequency in the control and treated groups.A common occurence in rodents maintained on ground feed is malocclusion. No parental females (F0) aborted or had physical or behavioral abnormalities during F1 gestation and lactation.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Survival was 100% for all groups of males and females at termination of the F0 generation animals except for the high-dose males where it was 97%. One male in this group (Animal No. C25201) was sacrificed at Week 10 in moribund condition due to an apparent fracture of the nasal bones.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Dose levels of 0.5% and 1.0% caused a reduction in body weight for the F0 males and the values were statistically significant at Weeks 9, 10, 18 and 19 for the 1.0% dose level and at Week 17 for the 0.5% dose level. No effect on body weight was seen for females at any dose level during the premating phase or during gestation. Although body weights for females during lactation were slightly lower than those of controls, the differences were not statistically significant.
There were no consistent significant differences in body weight gains for males. Body weight gains for 1.0% dose level females during Gestation Days 14 to 20 and 0 to 20 were significantly lower than those of controls .
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Although food consumptions for treated males were slightly lower than those of controls throughout the premating phase, these differences were significant only during Week 4 for the 0.5% dose level and during Weeks 4 and 5 for the 1.0% dose level. Food consumptions for 1.0% dose level females were lower than those of controls throughout lactation and statistically significant for all intervals except Days 10 through 14 and Days 17 through 21.
Compound consumption across groups was approximately proportional to the level of DSS administered in the test diets. As one would expect, it was decreased during the premating growth phase for both sexes(although to a greater extent in males ), remained low during gestation, and then increased appreciably during lactation.
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
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
There were no significant differences from the control group for male fertility indices during breeding for F1 litters. Fertility in males was defined by a female giving birth to a litter.
No significant differences from controls were observed for mating, fertility, or gestation indices, days required to mate, or the length of gestation. No parental females aborted or had physical or behavioral abnormalities during F1 gestation and lactation.
BODY WEIGHT (PARENTAL ANIMALS)

Table 1.Parental Body weights
Dose: 0 0.1% 0.5% 1%
Males Generation
Premating (age)
- Initial F0 (7 weeks) 232 238 241 238
F1 (6 weeks) 149 156 144 131*
F2 (7 weeks) 206 217 197 180
- Final F0 506 507 493 479
F1 510 512 492 447*
F2 531 536 492* 467*
Females
Premating
- Initial F0 (7 weeks) 165 163 165 166
F1 (6 weeks) 127 129 121 114*
F2 (7 weeks) 160 162 148 145
- Final F0 206 204 204 206
F1 281 296 271 255*
F2 285 290 271 269*
Gestation
- Initial F0 218 217 216 218
F1 278 294 267 258*
F2 288 291 277 270
- Final F0 361 365 360 350
F1 396 107 379 369*
F2 401 405 392 378*
Lactation
- Initial F0 280 280 275 267
F1 309 319 292 283*
F2 320 317 305 283*
- Final F0 288 294 290 273
F1 304 319 302 293
F2 313 316 314 300
* p < 0.05 (One-way ANOVA variance)
Dose descriptor:
NOAEC
Effect level:
0.1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Sex:
male/female
Basis for effect level:
body weight and weight gain
Remarks on result:
other: Generation: maternal/paternal toxicity
Key result
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Sex:
male/female
Basis for effect level:
other: no effects up to highest concentration
Remarks on result:
other: reproduction/offspring
Clinical signs:
no effects observed
Description (incidence and severity):
There were no treatment-related clinical observations for F1 males and females. Alopecia was noted with similar frequency in the control and treated groups .
As in the F0 generation, there were several instances of malocclusion and/or related signs. When necessary, the incisors of affected animals were clipped to alleviate the condition.
No parental females (F1) aborted or had physical or behavioral abnormalities during F2 gestation and lactation.

Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
Survival was 100% for all groups of males and females at termination of the F1 generation animals.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weights for F1 males and females at the 1.0% dose levels were significantly lower than those of controls throughout the premating phase, excluding Week 2 for females, and throughout the treatment period for males. Body weights for F1 males and females in the 0.5%. dose group were also low (although not statistically significant) during the entire premating phase.
Gestation body weights tor females were significantly lower than those of controls at the1.0% dose level throughout gestation. Body weights during lactation were significantly lower on days 0, 7, and 14 at the 1.0% dose level; body weights on Lactation Day 21 at the 1.0% dose level were also lower than those of controls, although this difference was not statistically significant.
Body weight gains were significantly lower than those of controls during Weeks 5 and 6 for males and during Weeks 3, 5, and 8 for females at the 1.0% dose level, and during Weeks 5 and 10 for females at the 0.5% dose level.

Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumptions were significantly lower than those of controls throughout the premating phase· (except f or Week 3) for males at the 1.0% dose level, and during Weeks 6 through 9 for males at the 0.5% dose level. Female food consumptions were significantly lower than those of controls during Weeks 7 and 8 at both the 0.5% and 1.0% dose levels.
Food consumptions during gestation were significantly greater than those of control only during Days 0 through 4 at the 0.1% dose level. Food consumptions during lactation were significantly lower during Days 7 through 10 at the 0.5% dose level, and during Days 4 through 7, 7 through 10, and 14 through 17 at the 1.0% dose level.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no treatment-related macroscopic observations in any animals examined in the study (F0, F1 and F2 adults and F3 weanlings).
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
There were no significant differences from the control group for male fertility indices during breeding for F2 litters .
No significant differences from controls were observed for mating, fertility, or gestation indices, days required to mate, or the length of gestation. No parental females aborted or had physical or behavioral abnormalities during F2 gestation and lactation.

Dose descriptor:
NOAEC
Effect level:
0.1 other: %
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
body weight and weight gain
Remarks on result:
other: maternal/paternal toxicity
Key result
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
other: no effects up to highest concentration
Remarks on result:
other: reproduction/offspring
Clinical signs:
no effects observed
Description (incidence and severity):
No significant differences from controls were observed for the total and mean number of pups born alive, litter size, survivability, or sex ratio. No pups from F1 litters had external abnormalities.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
No significant differences from controls were observed for the total and mean number of pups born alive, litter size, survivability, or sex ratio.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Male and female pup weights on days 4, 7, 14, and 21 and pup weight gains from Days 0 through 4, 4 through 7, 7 through 14, and 14 through 21 at the 1.0% dose level were significantly lower than those of controls. Male and female pup weights on day 21 and weight gains from Days 7 through 14 (females only), and from days 14 through 21 (males and females) for the 0.5% dose level were significantly lower than those of controls.
Food consumption and compound intake (if feeding study):
not examined
Description (incidence and severity):
Pups were allowed to nurse for 21 days before weaning. Food consumption after weaning is described under the parental generation.
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
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No pups from F1 litters had external abnormalities.
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
BODY WEIGHT (OFFSPRING)

Table 2.Offspring Body weights
Dose: 0 0.1% 0.5% 1%
Males
- Day 0 F1 6.5 6.8 6.4 6.6
F2 6.5 6.7 6.4 6.3
F3 6.7 6.7 6.8 6.1*
- Day 21 F1 15.6 14.7 13.7* 11.5*
F2 17.7 17.8 14.8* 12.4*
F3 19.7 19.9 17.6 13.4*
Females
- Day 0 F1 6.2 6.4 6.1 6.2
F2 6.1 6.4 6.1 6.0
F3 6.4 6.4 6.4 5.8*
- Day 21 F1 15.4 14.74 13.1* 10.8*
F2 16.5 17.1 14.5 11.4*
F3 18.6 19.3 16.0* 12.9*
* p < 0.05 (One-way ANOVA variance)
Dose descriptor:
NOAEC
Generation:
F1
Effect level:
0.1 other: %
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
body weight and weight gain
Clinical signs:
no effects observed
Description (incidence and severity):
No significant differences from controls were observed for the total and mean number of pups born alive, litter size, survivability, or sex ratio.
Alopecia, tremors, and rough hair coat were recorded on day 21 for three pups (two males and one female) from one F2 litter at the 0.5% dose level.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Description (incidence and severity):
No significant differences from controls were observed for the total and mean number of pups born alive, litter size, survivability, or sex ratio.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Beginning on day 7, male and female pup weights and pup weight gains at the 0.5% and 1.0% dose levels were consistently lower than those of controls; the differences were statistically significant for all values except day 7 female pup weight, male and female pup weight gains for the 0.5% dose level. On Day 4 of lactation no milk was observed in the abdomen of three pups from two litters in the control group, seven pups from five litters in the 0.1% dose group, 18 pups from 10 litters in the 0.5% dose group, and 10 pups from five litters in the 1.0% dose group.
Food consumption and compound intake (if feeding study):
not examined
Description (incidence and severity):
Pups were allowed to nurse for 21 days before weaning. Food consumption after weaning is described under the parental generation.
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
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Alopecia, tremors, and rough hair coat were recorded on day 21 for three pups (two males and one female) from one litter at the 0.5% dose level.
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
See under Details on results (F1).
Dose descriptor:
NOAEC
Generation:
F2
Effect level:
0.1 other: %
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
body weight and weight gain
Key result
Reproductive effects observed:
no
Lowest effective dose / conc.:
1 other: %

Extension on the F2 parents to generate an F3 litter generation:

Male fertility indices at the 0.5% and 1.0% dose levels during breeding for F3 litters were significantly higher than that of the control group.

No significant differences from controls were observed for mating, fertility, or gestation indices, days required to mate, or the length of gestation. No parental females aborted or had physical or behavioral abnormalities during F3 gestation and lactation.

Body weights for males were significantly lower than those of controls throughout the treatment period at the 1.0%; dose level, and from Week 3 through the duration of the treatment period for males at t he 0.5%; dose level. Body weights were significantly lower than those of controls for F2 females at the 1.0% dose level throughout the premating phase (except Weeks 0 and 2} , and during Weeks 1, 3 through 6, and 8 for females at the 0.5% dose level.

Gestation body weights were significantly lower than those of controls at the 1.0% dose level throughout gestation. Body weights during lactation were significantly lower on Days 0, 7, and 14 at the 1.0% dose level; mean body weight on lactation Day 21 at the 1.0% dose level also was lower than that of the control, although this difference was not statistically significant.

Body weight gains were significantly lower than these of controls during weeks 4, 5 through 8, 13, 15, 17 , 20, 23, and 24 formales at the 1.0% dose level , during Weeks 4, 6, 7, 13, 16, 23, and 24 for males at the 0.5% dose level, and during Week 3 for males at the 0.1% dose level. There were no consistent significant differences in weight gains for females during the treatment period.

Body weight gains during lactation were significantly higher than these of the control between Days 14 and 21 at the 1.0% dose level, and between Days 0 and 21 for the 0.5% and 1.0% dose levels.

Food consumptions were significantly lower than those of controls during Weeks 2, 5 through 7, and 10 for males at the 1.0% dose level, and during Weeks 5 through 7 for males at the 0.5% dose level, Female food consumptions were significantly lower than those of controls during Week 2 at both the 0.5% and 1.0% dose levels, and were significantly higher during Week 7 at the 0.1% dose level.

F3 Litter data:

No significant differences from controls were observed for the mean number of pups born alive, litter size, survivability, or sex ratio. The proportions of the total number of pups born alive and found dead at the 1.0% dose level were significantly lower and higher, respectively, than those of the controls. These differences may merely reflect the effect of a slight reduction in the number of litters in the control group, and a disproportionate influence of very few litters in the high-dose group; i.e. , two litters accounted for 13 (six plus seven) of the 17 pups found dead; there was one dead pup each in four other litters. Male and female pup weights and pup weight gains at the 1.0% dose level were significantly lower than these of controls throughout lactation. At the 0.5% dose level, pup weight gains for males and females from Days 4 through 7, for males from Days 7 through 14, and tor fema1es from Days 14 through 21 were significantly lower than these of the control, as were male pup weights on Day 14 and male and female pup weights on Day 21. One pup in the0.5% dose group and 17 pups from seven litters In the 1.0% dose group had no milk in the abdomen on Day 4 of lactation. One female pup in the control group was icteric on Day 4. Another control pup from a different litter was missing the left eye. Five males and five females from one litter at the 1.0% dose level had urine stains on Day 21.

Conclusions:
Read-across substance DSS administered in the diet to three successive generations of rats at levels of 0.5% and 1.0% caused a reduction in body weights for parental males of all generations and for F1 and F2 females. Pup weights at the 0.5% and 1.0% dose levels were lower than those of the control in all three generations. However, the reduced body weight did not interfere with growth and development or reproductive performance, and the test substance had no adverse effects at levels on the reproductive function of either sex in any generation up to 1% in the diet.
Based on the results of this study, when DSS was administered in the diet to three succesive generations of rats, the no-observable-effect level (NOEL) for body weights of parental animals and offspring was 0.1%; the NOEL for reproductive parameters was 1.0% DSS.
Executive summary:

Read-across substance Docusate sodium was administered in the diet to three successive generations at levels of 0.1%, 0.5% and 1.0% in the diets of 30 males and 30 females per group, dosed for 10 and 2 weeks respectively. The dose levels of 0.5% and 1.0% caused a reduction in body weights for parental males of all generations and for F1 and F2 females. Pup weights at the 0.5% and 1.0% dose levels were lower than those of the control in all three generations. However, the reduced body weight did not interfere with growth and development or reproductive performance, and the test substance had no adverse effects at levels on the reproductive function of either sex in any generation up to 1.0% in the diet. There were no other effects on parental or reproductive parameters.

Based on the results of this study, when DSS was administered in the diet to three succesive generations of rats, the no-observable-effect level (NOEL) for body weights of parental animals and offspring was 0.1%; the NOEL for reproductive parameters was 1.0% DSS.

Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Justification for type of information:
Read across argumentation is provided in Section 13.
Reason / purpose:
read-across source
Clinical signs:
no effects observed
Description (incidence and severity):
The overall appearance of the animals fed AEROSOL OT was good, except for an occasional outbreak of respiratory distress. This was characterized by raspy breathing and occasional red-colored nasal exudates. No other conditions were noted.
Dermal irritation (if dermal study):
not examined
Mortality:
not specified
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Males fed AEROSOL OT had mean body weights (prior to the paring for mating) about 7 – 8% less than that of control males until the F2 generation. In that generation the 0.5% males were essentially similar to control males in body weight, while the 1% males weighed about 5% less than the controls. All groups of females had essentially similar body weights at any given mating period.

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
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
F0 first mating: Fertility, Gestation, and Viability Indices are not too dissimilar for the 3 groups of animals, and they could be interpreted as reflecting the normal variability which is encountered in studies of this nature. The Lactation Index is down for each group. Of the pups alive at 5 days, less than two-thirds of the controls and less than one-half of the test groups survived to weaning. The failure of substantial numbers of pups to survive beyond 5 days was puzzling. The low Lactation Indices for the 2 test groups undoubtedly reflect, in part, whatever unknown etiological factors were present that contributed to the relatively poor performance of the control group in this index. However, since both test groups showed greater decreases in the Lactation Index, and since this index was almost identical for both test groups, it was felt that there could be an additional cause for the depression of the index in the test groups. Since Aerosol OT has a bitter taste, it was surmised that if the material was secreted into the maternal milk, it might affect palatability and the subsequent acceptance of the milk by the pups. This could account for the failure of the pups to be weaned despite their good survival in the first post-natal days. Consequently, the F0 generation animals were remated.
F0 second mating: Fertility and Gestation Indices are quite high and comparable for the test and control groups. While one or 2 animals in each test group failed to conceive, the only pregnancy that failed to go to term was in a control animals.
Viability of test pups was slightly less than that of the controls, but this clearly was not dose-related in that viability of the 1% group was greater than that of the 0.5% group. Relative to the first mating, the Lactation Index was increased for each group of pups. The value of 66 for the 1% group is, in fact, greater than the Lactation Index observed in control rats for the first mating. Of more importance was the fact that 75% of the test dams weaned one or more pups in the second mating, compared to 55% in the first mating. Consequently, an adequate number of pups from a representative number of litters was available to continue the study.
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Sex:
male/female
Basis for effect level:
other: general toxicity
Dose descriptor:
NOAEL
Effect level:
>= 790 - <= 890 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Remarks:
in the diet
Sex:
male/female
Basis for effect level:
other: general toxicity
Clinical signs:
not specified
Dermal irritation (if dermal study):
not examined
Mortality:
not specified
Body weight and weight changes:
no effects observed
Description (incidence and severity):
At weaning mean body weight for each group was almost identical.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
The 3 parameters which had shown an apparent decrease in the second mating of the F0 generation (Viability and Lactation Indices and mean body weight at weaning) were not affected in this phase of the study.
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Basis for effect level:
other: no general and reproductive toxicity
Clinical signs:
not specified
Dermal irritation (if dermal study):
not examined
Mortality / viability:
not specified
Body weight and weight changes:
not examined
Description (incidence and severity):
only weights at weaning
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
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Dose descriptor:
NOAEC
Generation:
F1
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Sex:
male/female
Basis for effect level:
other: No litter and fetal toxicity
Clinical signs:
not specified
Dermal irritation (if dermal study):
not examined
Mortality / viability:
not specified
Body weight and weight changes:
not examined
Description (incidence and severity):
only weights at weaning
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
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Dose descriptor:
NOAEC
Generation:
F2
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Sex:
male/female
Basis for effect level:
other: No litter and fetal toxicity
Reproductive effects observed:
not specified

All of the pups up to the first mating of the second generation (F1a, F1b, F2) and those from the second mating of the second generation (F3b pups) were observed for gross defects and discarded without autopsy.

Autopsies were performed on all animals produced by the first mating of the second generation (F3a pups) when they were weaned at 21-22 days of age. Pups which died prior to weaning were autopsied immediately after death unless postmortem autolysis made the results valueless.

Microscopic study of tissues showed developmental, parasitic, and rarely, infectious lesions which were similar In all groups. There was no histopathological evidence of injury attributable to administration of the test article. Skeletal abnormalities were either of a traumatic nature, probably caused by the handling and trimming involved in processing the skeletons, or the presence of an extra sternebrae in the sternum between the 5th and 6th sternebrae. Whether the latter was actually an accessory, or merely a center of ossification for number 5 or 6 which had not yet fused with the corresponding sternebrae, cannot be stated. The infrequent occurrence of this structure makes it likely a truly accessory sternebrae. Consequently, none of the skeletal changes seen were attributed to parental exposure of test material.

It is concluded that feeding of test material to rats from weaning through reproductive age for successive generations at levels of 1%, or less, did not produce lesions or anomalies in the offspring which could be attributed to the compound.

Conclusions:
It is concluded that feeding of read-across substance AEROSOL OT to rats from weaning through reproductive age for succesive generations at levels of 1% or less, did not produce lesions or anomalies in the offspring which could be attributed to the compound.
Executive summary:

Young albino rats were given 0.5% and 1% read-across substance AEROSOL OT and they were mated at the age of 3 - 4 months (one male and one female). In the first mating of the F0 generation and the second mating of the F2 generation, pups were weaned directly onto the diets which were being fed to their parents. In the other 3 matings of this study, dams were given a control diet on the day before they were expected to cast their litters. When they were weaned, pups of these 3 litters were given diets containing the same levels of test material that their progenitors had been receiving. When pups were 3-4 months of age, they were in turn mated, and the process was repeated with their pups for a total of 3 generations.

The first mating of the F0 generation and the second mating of the F2 generation were similar in that the dams were continuously fed and the pups were weaned directly onto diets containing test material. In both of these matings, the fertility and gestation indices were high and comparable. The viability index was good, albeit slightly down for the F3b pups, while the lactation index was depressed for both of these matings. In addition, the mean weight of the pups at weaning decreased with increasing concentrations of test material in the diet of the dams.

For the other 3 matings (F1b, F2, F3a pups), test material was removed from the diet of the dam on the day before they were expected to cast their litters. After weaning, the pups were placed on a control or dosed diets. In the second mating of the F0 animals, the viability and lactation indices and the mean weight of the test pups at weaning still showed decreases relative to the control values. However, in the 2 subsequent matings, all indices for the dosed animals were numerically high and compared favorably with the corresponding control values, Also, the mean weight of the pups at weaning was essentially similar fox all groups. Consequently, it is concluded that diets containing 1% or less had no adverse effect on the reproduction and lactation performance of rats. The lowering of the survival rate and the mean body weight of the F1a and F3b pups is attributed to an impairment of nutrition as a result of the taste which is believed to have been secreted into the milk of the dams.

Pups of all litters, including those which died before weaning, were examined for gross defects. Autopsies were performed, however, only on pups from the first mating of the F2 animals. Those pups were killed at weaning. Immediately after death, the 2 males and 2 females which were the smallest or appeared least healthy of each litter were set aside, while the others were autopsied. Portions of all major organs were taken for histopathology processing and examination from one male and female from each litter. The other male and female were skinned and eviscerated, and the carcasses cleared, and the skeletons stained and examined for defects.

Microscopic study of tissues showed developmental, parasitic, and rarely, infectious lesions which were similar In all groups. There was no histopathological evidence of injury attributable to administration of the test article. Skeletal abnormalities were either of a traumatic nature, probably caused by the handling and trimming involved in processing the skeletons, or the presence of an extra sternebrae in the sternum between the 5th and 6th sternebrae. Whether the latter was actually an accessory, or merely a center of ossification for number 5 or 6 which had not yet fused with the corresponding sternebrae, cannot be stated. The infrequent occurrence of this structure makes it likely a truly accessory sternebrae. Consequently, none of the skeletal changes seen were attributed to parental exposure of test material.

It is concluded that feeding of test material to rats from weaning through reproductive age for successive generations at levels of 1%, or less, did not produce lesions or anomalies in the offspring which could be attributed to the compound.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
750 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Klimisch 2
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No test data were available for current substance, however read across data were available from Docusate sodium (CAS 577 -11 -7). Justification for read across within the category of Di-ester sulphosuccinates is documented in a separate document attached in Section 13.

Multigeneration studies

-A key study for reproductive toxicity was available for read across substance Docusate sodium as a 3-generation toxicity study at dietary dose levels of 0.1, 0.5 and 1.0% in the diet (MacKenzie, 1986). The study was conducted according to OECD 416 and GLP guidelines, and was considered to be reliable, adequate and relevant. The test substance caused a reduction in body weights at the dose levels of 0.5 and 1% in the diet for parental males of all generations and for F1 and F2 females. Pup weights at the 0.5% and 1.0% dose levels were lower than those of the control in all three generations. However, the reduced body weight did not interfere with growth and development or reproductive performance, and the test substance had no adverse effects at levels on the reproductive function of either sex in any generation up to 1%. There were no other effects on parental or reproductive parameters. Based on the results of this study, when Docusate sodium was administered in the diet to three successive generations of rats, the no-observable-effect level (NOEL) for body weights of parental animals and offspring was 0.1%; the NOEL for reproductive parameters was 1.0% Docusate sodium, which was considered to correspond with approximately 750 mg/kg bw/day.

- In a supporting successive 2-generation toxicity study in rats, dietary doses of Docusate sodium given were 0.5 and 1% (Levinskas & Shaffer, 1970). In the first mating of the F0 generation and the second mating of the F2 generation, pups were weaned directly onto the diets which were being fed to their parents. In the other 3 matings of this study, dams were given a control diet on the day before they were expected to cast their litters to avoid a bitter taste of the milk. When they were weaned, pups of these 3 litters were given diets containing the same levels of test material that their progenitors had been receiving. When pups were 3-4 months of age, they were in turn mated, and the process was repeated with their pups for a total of 3 generations. Pups of all litters, including those which died before weaning, were examined for gross defects. Autopsies were performed, however, only on pups from the first mating of the F2 animals. Those pups were killed at weaning. Immediately after death, the 2 males and 2 females which were the smallest or appeared least healthy of each litter were set aside, while the others were autopsied. Portions of all major organs were taken for histopathology processing and examination from one male and female from each litter. The other male and female were skinned and eviscerated, and the carcasses cleared, and the skeletons stained and examined for defects. In both the first mating of the F0 generation and the second mating of the F2 generation, the fertility and gestation indices were high and comparable. The viability index was good, albeit slightly down for the F3b pups, while the lactation index was depressed for both of these matings. In addition, the mean weight of the pups at weaning decreased with increasing concentrations of test material in the diet of the dams. In the second mating of the F0 animals, the viability and lactation indices and the mean weight of the test pups at weaning still showed decreases relative to the control values. However, in the 2 subsequent matings, all indices for the dosed animals were numerically high and compared favorably with the corresponding control values. Also, the mean weight of the pups at weaning was essentially similar for all groups. Consequently, it is concluded that diets containing 1% or less had no adverse effect on the reproduction and lactation performance of rats. The lowering of the survival rate and the mean body weight of the F1a and F3b pups is attributed to an impairment of nutrition as a result of the taste which is believed to have been secreted into the milk of the dams.

Microscopic study of tissues showed findings which were similar in all groups. In processing the skeletons,the presence of an extra sternebrae in the sternum between the 5th and 6th sternebrae was not considered to parental exposure of test material.

It is concluded that feeding of test material to rats from weaning through reproductive age for successive generations at levels of 1%, or less, did not produce lesions or anomalies in the offspring which could be attributed to the compound.

Conclusion

Multigeneration studies with read across substance Docusate dodium (CAS 577-11-7) showed slight maternal/paternal toxicity at 0.5 and 1% in the diet, however this was not confirmed in the second study. The test article was considered to have influenced the taste of the milk for which adaptations were done in the second study. From both studies, it can be concluded that the substance up to 1% in the diet did not lead to effects on fertility or postnatal development; this concentration corresponds with 750 mg/kg bw/day, which is higher than the NOAEL for paternal/maternal toxicity.
Based on the absence of reproductive findings in the repeated dose studies and the multigeneration studies no further testing is needed.


Effects on developmental toxicity

Description of key information
Prenatal developmental toxicity was tested by dietary administration of read across substance Docusate sodium in rats from day 6 to 15 of gestation. 1% in the diet was a maternal and developmental NOAEL, whereas at 2% in the diet visceral and skeletal anomalies were observed, which was secondary to maternal toxicity. This was confirmed in a similar study with Docusate calcium given at subtoxic and toxic dose levels, where the same could be observed.  
Based on the absence of developmental findings in the teratogenicity study, no further testing is needed.
Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
Read across argumentation is provided in Section 13.
Reason / purpose:
read-across source
Strain:
Abyssinian
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Among rats given dietary levels of 2.0% DSS, there were significant depressions in maternal weight-gains.
Rats fed diets containing 1.0% level of DSS showed no significant maternal effects on the various parameters.
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:
not specified
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Details on results:
Rats fed diets containing 1.0% level of DSS) showed no significant maternal effects on the various parameters. Among rats given dietary levels of 2.0% DSS, there were significant depressions in maternal weight gains.
Number of abortions:
no effects observed
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
effects observed, treatment-related
Description (incidence and severity):
In the 2.0% DSS group 1 pregnancy with total resorptions was observed (No statistical significance). No pregnancy with total resorptions was observed in the control or 1.0% DSS group.
Early or late resorptions:
effects observed, treatment-related
Description (incidence and severity):
Among rats given dietary levels of 2.0% DSS, there were significant increases in the number of resorptions of 13.7% as compared to the control frequency of 5.6%.
Dead fetuses:
no effects observed
Description (incidence and severity):
0.5% occurrence of dead fetuses was seen in the 2.0% DSS group versus 0.7% in the control group. No dead fetuses were observed in the 1.0% DSS group.
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:
not examined
Details on maternal toxic effects:
Maternal toxic effects:yes. Remark: 2.0% in the diet
Key result
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Basis for effect level:
body weight and weight gain
early or late resorptions
Key result
Dose descriptor:
NOAEL
Effect level:
1 074 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
body weight and weight gain
early or late resorptions
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): There is no postnatal evaluation in an OECD 414 study.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
There is no significant reduction in viable fetuses in the dosed animals animals compared to control animals.
Changes in sex ratio:
not specified
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
Description (incidence and severity):
There is no postnatal evaluation in an OECD 414 study.
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Near toxic or toxic dietary levels of 2.0% DSS produced significant incidences of gross abnormalities either among litters (25.0%) or fetal populations (20.2%) as compared to none in the controls. These abnormalities consisted of cranial buble, exencephaly, spina bifida (not significant), microphtalmia or anophtalmia (not significant).
Skeletal malformations:
no effects observed
Visceral malformations:
effects observed, treatment-related
Description (incidence and severity):
The visceral observations confirmed the significance of the exencephalous characteristics and anophtalmia for the group given dietary levels of 2.0% DSS.
Other effects:
effects observed, treatment-related
Description (incidence and severity):
In the 2.0% DSS group, skeletal observations revealed a significant incidence of variations including incomplete ossification to absence of the various cranial bones, a curved or open vertebral column, and a variety of defects of the vertebrae and ribs.
Details on embryotoxic / teratogenic effects:
Details on embryotoxic / teratogenic effects:
See Table 1-4.
Key result
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
external malformations
visceral malformations
other:
Remarks on result:
other: secondary to high maternally toxic dose
Key result
Dose descriptor:
NOAEL
Effect level:
1 074 mg/kg bw/day
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
external malformations
visceral malformations
other: skeletal variations
Abnormalities:
effects observed, treatment-related
Localisation:
external: cranium
skeletal: skull
skeletal: rib
visceral/soft tissue: central nervous system
visceral/soft tissue: eye
Description (incidence and severity):
only at 2.0% dietary level.
Developmental effects observed:
yes
Lowest effective dose / conc.:
2 other: %
Treatment related:
yes
Relation to maternal toxicity:
developmental effects as a secondary non-specific consequence of maternal toxicity effects
Dose response relationship:
yes
Relevant for humans:
no

Table 1. Maternal and fetal results of pregnant rats given various amounts if DSS in their diets during  gestational days 6 through 15.

Parameter

 Control        

1.0% DSS

2.0% DSS

Maternal

Group  (I-A)

(II-A)

(II-B)

No. of pregnant rats

43

22

20

No. of pregnancies with total resorptions

0

0

1

No. of pregnancies with viable fetuses

43

22

19

Average weight gain of dams with viable fetuses(g):

 

 

 

Days 6 to 15

78

86

52*

Days 15 to 21

66

67

77

Average, apparent food intake of dams with viable fetuses (g/rat/day):

 

 

 

Days 6 to 15

22.5

24.8

21.4

Days 15 to 21

28.6

32.1

33.4

Calculated compound consumed (mg/kg/day)

--

1074

1988

Litters

 

 

 

Total number of:

implantations

 

411

 

203

 

219

Resorptions

(% occurence)

23

(5.6)

8

(3.9)

30*a

(13.7)

Dead fetuses

(% occurrence)

3

(0.7)

0

1

(0.5)

Viable fetuses

(% occurrence)

385

(93.7)

195

(96.1)

188

(85.5)

Fetal weight (g)

4.6

5.2

4.7

Litters size (viable fetuses)

8.9

8.9

9.9

External major malformations1:

No. of litters affected

(% occurrence)

 

 

0

 

 

0

 

 

5*

(25.0)

No. of fetuses affected

(% occurrence)

 

0

 

0

36*a

(20.2)

* Significantly different from control (p< 0.05)

a Significance by Chi-square, but not Mann-Whitney U test

1 Primarily, exencephaly varying degrees and associated anomalies (See Table 2)

    

Table 2. Morphological observations of fetuses delivered from rats given DSS in their diets on gestational days 6 through 15.

Morphology

 Control

1.0% DSS

2.0% DSS

External observations1:

Group (I-A)

(II-A)

(II-B)

Total number examined

388a

195

189

Major anomalies:

  Adactyly

 

0

 

0

 

0

  Hemimelia

0

0

0

  Schistocelia

0

0

2

  Dome shaped head

0

0

0

  Cranial bubble (1-2mm)

0

0

9*

  Exencephaly

0

0

18*

  Exencephaly (cleft condition)

0

0

7*

  Anencephaly

0

0

0

  Spina bifida

0

0

6

  Macroglossia

0

0

0

  Micro- or anophtalmia

0

0

3

Defects:

  Hematoma (subcutaneous)

 

2

 

0

 

0

  Edamatous abdomen

0

0

0

  Tail short & curled

0

0

0

  Abducted fifth digit, left

   Rear foot

0

0

1

1 Fetuses may have more than one defect

a Fifty-four fetuses examined grossly only. (Shipment c valid as controls only)

      *Significantly different from control (p< 0.05) by Chi-square only

 

Table 3. Visceral observations of fetuses delivered from rats given DSS in their diets on gestation days  6 through 15.

Visceral observations

Dose:      Control

1.0 % DSS

2.0% DSS

Groups:       (I-A)

(II-A)

(II-B)

Total number of fetuses examined

165a

98

91

Defects1:

  Exencephalous   characteristics                     

 

0

 

0

 

11*

  Dilated lateral ventricles

1

3

5

  Microphtalmia

0

1

0

  Anolphtalmia

0

0

23*

  Retinal foldings

0

0

0

  Anotia or microtia

0

0

0

  Cleft palate

0

0

1

  Situs transversus – aorta, esophagus

  & stomach

1

0

0

  Intestinal agenesis

0

0

0

  Arch of aorta absent or right sided

0

0

0

  Diaphragmic hernia

0

0

1

  Dilated renal pelves

2

0

3

  Ectopic kidneys(s) &/or variation in size

1

0

0

  Renal agenesis

0

0

2

  Dilated ureters

6

0

3

  Adrenal agenesis

0

0

1

  Testes – ectopic or enlarged

1

0

1

  Hermaphroditism

0

0

3

1Fetuses may have more than one defect

aExcludes 1 fetus lost

*Significantly different from control (p<0.05) by Chi-square only

Table 4. Skeletal observations of fetuses delivered from rats given DSS in their diets on gestation days  6 through 15.

 

Skeletal observations

Dose:      Control

1.0 % DSS

2.0% DSS

Group  (I-A)

(II-A)

(II-B)

Total number of fetuses examined

167a

97

98

Defects1:

  Cranial bones,

  incomplete to lack of ossification :

   Nasal                    

 

 

 

0

 

 

 

0

 

 

 

4

   Frontal

1

0

20*

   Parietal

1

1

19*

   Interparietal

1

2

18*

   Supraoccipital

0

0

15*

   Exoccipital

0

0

2

   Atlas

0

0

1

   Zygomatic

0

0

1

   Premaxilla

0

0

1

   Tympanic bullae

0

0

5

   Mandibles

0

0

1

   Hyoid

0

0

3

  Eye orbit, reduction

0

0

0

  Exoccipital, fused to atlas

0

0

0

  Vertebrla column, curved &/or open

0

0

5

  Vertebrae:

 

 

 

   misshapened &/or retarded 

   development

0

0

5

   thoracic, bipartite centra

2

1

5

   lumbar, bipartite centra

0

0

2

  Sternebrae:

 

 

 

   fused

0

0

0

   hypoplastic to absent

0

0

1

   one or two absent

1

0

0

   staircase

0

0

3

   bipartite

0

0

2

  Rib(s):

 

 

 

   accesory

6

5

5

   Absent or less developed

0

0

7*

   wavy

2

2

0

   fused

0

0

2

  Pelvic, hypoplastic to absent

0

0

0

  Brachydactyly

0

0

0

  Syndactyly

0

0

0

  Adactyly

0

0

0

  Hemimelia & small scapula

0

0

 0

1Fetuses may have more than one defect

aExcludes 1 fetus destroyed during cleaning process

*Significantly different from control (p<0.05) by Chi-square only

 

Conclusions:
Subtoxic dietary levels of 1.0% read-across substance docusate sodium ingested on gestational days 6 through 15 showed no adverse effects on the various maternal or fetal parameters. Near toxic or toxic dietary levels of 2.0% DSS produced significant incidences of resorptions (13.7%) and gross abnormalities either among litters (25.0%) or fetal populations (20.2%) as compared to controls. Interpretation of the results of the present experiments, in which only maternally toxic dose levels induce teratogenicity, indicates no real hazard with the recommended human use of these surfactants.
Executive summary:

Prenatal developmental toxicity was studied in rats dosed from day 6 to day 15 of gestation by dietary administration of read-across substance docusate sodium at dose levels of 1.0 and 2.0 % in the diet. Subtoxic dietary levels of 1.0% showed no adverse effects on the various maternal or fetal parameters. Near toxic or toxic dietary levels of 2.0% docusate sodium produced significant depressions in maternal weight-gains and increased incidences of resorptions (13.7%) and gross abnormalities either among litters (25.0%) or fetal populations (20.2%) as compared the controls. These abnormalities consisted primarily of exencephaly of varying degrees with, at times, spina bifida, anophtalmia and associated skeletal defects. The visceral observations confirmed the significance of the exencephalous characteristics and anophtalmia for the group given dietary levels of 2.0%. In this group, skeletal observations revealed a significant incidence of incomplete ossification to absence of the various cranial bones, a curved or open vertebral column, and a variety of defects of the vertebrae and ribs. There were significant depressions in maternal weight gains in the 2.0% DSS-group. Interpretation of the results of the present experiment, in which only maternally toxic dose levels induce teratogenicity, indicates no real hazard with the recommended human use of these surfactants.

The concentration of 1% in the diet is considered as maternal and developmental NOAEL. This dose level corresponded with 1074 mg/kg body weight, as calculated in the study.

Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reason / purpose:
read-across source
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Among rats given dietary levels of 2.0% DCS, there were significant depressions in maternal weight gains.
Rats fed diets containing 1.5% , 1.0%, 0.5% level of DCS showed no significant maternal effects, however a non-significant decrease in weight gain was observed at 1.5% from day 6-15.
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:
not specified
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
effects observed, treatment-related
Description (incidence and severity):
In the 2.0% DCS group 1 pregnancy with total resorptions was observed (no statistical significance). No pregnancy with total resorptions was observed in the control or lower concentration DCS groups.
Early or late resorptions:
effects observed, treatment-related
Description (incidence and severity):
Among rats given dietary levels of 2.0% DCS, there were significant increases in the number of resorptions of 13.4% as compared to the control frequency of 5.6%.
Dead fetuses:
no effects observed
Description (incidence and severity):
0.3% occurrence of dead fetuses was seen in the 2.0% DCS group and 1.2% occurrence ion the 1.5% DCS group versus 0.7% in the control group. No dead fetuses were observed in the lower dose groups.
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:
not examined
Details on maternal toxic effects:
Maternal toxic effects:yes. Remark: 2.0% in the diet
Key result
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
in the diet
Basis for effect level:
body weight and weight gain
early or late resorptions
Key result
Dose descriptor:
NOAEL
Effect level:
1 060 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Remarks:
in the diet
Basis for effect level:
body weight and weight gain
early or late resorptions
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): There is no postnatal evaluation in an OECD 414 study.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
There is no significant reduction in viable fetuses in the dosed animals animals compared to control animals.
Changes in sex ratio:
not specified
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
Description (incidence and severity):
There is no postnatal evaluation in an OECD 414 study.
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Near toxic or toxic dietary levels of 1.5% and 2.0% DCS produced significant incidences of gross abnormalities either among litters ( 25.9% and 18.2% respictively) or fetauses (19.3% and 11% respectively) as compared to none in the controls. These abnormalities mainly consisted of dome shaped head, exencephaly, cranial bubble, spina bifida, macroglossia and microphtalmia or anophtalmia.
Skeletal malformations:
no effects observed
Visceral malformations:
effects observed, treatment-related
Description (incidence and severity):
The visceral observations confirmed the significance of the exencephalous characteristics and anophtalmia for the group given dietary levels of 1.5% and 2.0% DCS.
Other effects:
effects observed, treatment-related
Description (incidence and severity):
In the 1.5% and 2.0% DCS group, skeletal observations revealed a significant incidence of variations including incomplete ossification to absence of the various cranial bones, a curved or open vertebral column, and a variety of defects of the vertebrae and ribs.
Details on embryotoxic / teratogenic effects:
Details on embryotoxic / teratogenic effects:
See Table 1.
Key result
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
other: active ingredient diet
Sex:
male/female
Basis for effect level:
external malformations
visceral malformations
other: skeletal variations
Remarks on result:
other: secondary to high maternally toxic dose
Key result
Dose descriptor:
NOAEL
Effect level:
1 060 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
external malformations
visceral malformations
other: skeletal variations
Remarks on result:
other: secondary to high maternally toxic dose
Abnormalities:
effects observed, treatment-related
Localisation:
external: cranium
skeletal: skull
skeletal: rib
skeletal: vertebra
visceral/soft tissue: central nervous system
visceral/soft tissue: eye
Description (incidence and severity):
Only at 1.5% and 2.0% dietary levels (1611 and 2232 mg/kg bw/day actual dose active ingredient received, respectively).
Developmental effects observed:
yes
Lowest effective dose / conc.:
1.5 other: %
Treatment related:
yes
Relation to maternal toxicity:
developmental effects as a secondary non-specific consequence of maternal toxicity effects
Dose response relationship:
yes
Relevant for humans:
no

Table 1. Maternal and fetal results of pregnant rats given various amounts of DCS in their diets during  gestational days 6 through 15.

Parameter

Control        

0.5% DCS

1.0% DCS

1.5% DCS

2.0% DCS

Maternal

 

 

 

 

 

No. of pregnant rats

43

25

33

27

33

No. of pregnancies with total resorptions

0

0

0

0

1

No. of pregnancies with viable fetuses

43

25

33

27

32

Average weight gain of dams with viable fetuses(g):

 

 

 

 

 

Days 6 to 15

78

76

81

62

55*

Days 15 to 21

66

71

64

71

67

Avarage, apparent food intake of dams with viable fetuses (g/rat/day):

 

 

 

 

 

Days 6 to 15

22.5

22.8

23.8

23.5

24.7

Days 15 to 21

28.6

28.0

30.0

29.8

37.7

Calculated compound consumed (mg/kg/day)

--

499

1060

1611

2232

Litters

 

 

 

 

 

Total number of:

implantations

 

411

248

314

250

328

Resorptions

(% occurence)

23

(5.6)

12
(4.8)

25
(8.0)

44*
(3.9)

30*a

(13.7)

Dead fetuses

(% occurrence)

3

(0.7)

0

0

3
(1.2)

1
(0.3)

Viable fetuses

(% occurrence)

385

(93.7)

235

(95.2)

289

(92.0)

225

(90.0)

283

(86.3)

Fetal weight (g)

4.6

4.6

5.0

4.2

4.6

Litter size (viable fetuses)

8.9

9.4

8.8

8.3

8.8

External major malformations1:

No. of litters affected

(% occurrence)

 

 

0

 

 

1

 

 

0

 

 

7*
(25.9)

 

 

6*
(18.2)

No. of fetuses affected

(% occurrence)

 

0

 

1

 

0

 

44*a
(19.3

 

36*a

(11.0)

* Significantly different from control (p< 0.05)

a Significance by Chi-square, but not Mann-Whitney U test

1 Primarily, exencephaly varying degrees and associated anomalies (See TABLE 2)

   

 

Table2. Morphological observations of fetuses delivered from rats given DCS in their diets on gestational days 6 through 15.

Morphology

Control        

0.5% DCS

1.0% DCS

1.5% DCS

2.0% DCS

External observations1:

 

 

 

 

 

Total number examined

388a

236

289

228

284

Major anomalies:

  Adactyly

 

0

 

0

 

0

 

0

 

4

  Hemimelia

0

1

0

0

0

  Schistocelia

0

0

0

1

1

  Dome shaped head

0

0

0

6*

0

  Cranial bubble (1-2mm)

0

0

0

8*

3

  Exencephaly

0

0

0

23*

20*

  Exencephaly (cleft condition)

0

0

0

3

2

  Anencephaly

0

0

0

0

1

  Spina bifida

0

0

0

7*

14*

  Macroglossia

0

0

0

4

7*

  Micro- or anophtalmia

0

0

0

9*

4

Defects:

  Hematoma (subcutaneous)

 

2

 

1

 

1

 

4

 

3

  Edamatous abdomen

0

0

0

1

0

  Tail short & curled

0

0

0

1

0

  Abducted fifth digit, left

   Rear foot

0

0

0

0

0

1 Fetuses may have more than one defect

a Fifty-four fetuses examined grossly only. (Shipment c valid as controls only)

      *Significantly different from control (p< 0.05) by Chi-square only

Table 3. Visceral observations of fetuses delivered from rats given DCS in their diets on gestation days 6 through 15.

Visceral observations

Control        

0.5% DCS

1.0% DCS

1.5% DCS

2.0% DCS

 

 

 

 

 

Total number of fetuses examined

165

116

146

112

142

Defects1:

  Exencephalous   characteristics                     

 

0

 

0

 

0

 

15*

 

13*

  Dilated lateral ventricles

1

0

6

6

1

  Microphtalmia

0

0

0

2

0

  Anolphtalmia

0

0

0

19*

14*

  Retinal foldings

0

0

0

0

4

  Anotia or microtia

0

0

0

1

1

  Cleft palate

0

0

0

3

0

  Situs transversus – aorta, esophagus & stomach

1

0

0

0

0

  Intestinal agenesis

0

0

0

1

0

  Arch of aorta absent or right sided

0

0

0

2

0

  Diaphragmic hernia

0

0

0

0

0

  Dilated renal pelves

2

0

0

4

2

  Ectopic kidneys(s) &/or variation in size

1

0

1

0

1

  Renal agenesis

0

0

0

1

0

  Dilated ureters

6

10

1

21

5

  Adrenal agenesis

0

0

0

0

0

  Testes – ectopic or enlarged

1

0

0

2

1

  Hermaphroditism

0

0

0

1

1

1Fetuses may have more than one defect

*Significantly different from control (p<0.05) by Chi-square only

 

Table 4. Skeletal observations of fetuses delivered from rats given DCS in their diets on gestation days 6 through 15.

Skeletal observations

Control        

0.5% DCS

1.0% DCS

1.5% DCS

2.0% DCS

 

 

 

 

 

Total number of fetuses examined

167a

120

143

113

141

Defects1:

  Cranial bones,

  incomplete to lack of ossification :

   Nasal                    

 

 

 

0

 

 

 

0

 

 

 

0

 

 

 

5

 

 

 

4

   Frontal

1

0

0

17*

19*

   Parietal

1

0

2

18*

20*

   Interparietal

1

0

1

17*

17*

   Supraoccipital

0

0

4

14*

17*

   Exoccipital

0

0

0

0

0

   Atlas

0

0

0

0

0

   Zygomatic

0

0

0

1

0

   Premaxilla

0

0

0

0

2

   Tympanic bullae

0

0

0

0

0

   Mandibules

0

0

0

1

2

   Hyoid

0

0

0

0

7*

Eye orbit, reduction

0

0

0

3

1

Exoccipital, fused to atlas

0

0

0

0

1

Vertebrla column, curved &/or open

0

0

0

6*

6*

ertebrae:

 

 

 

 

 

   misshapened &/or retarded 

   development

0

0

0

3

9*

   thoracic, bipartite centra

2

1

2

7

14*

   lumbar, bipartite centra

0

0

0

4

5

Sternebrae:

 

 

 

 

 

   fused

0

0

0

0

2

   hypoplastic to absent

0

0

0

5

0

   one or two absent

1

1

0

5

0

   staircase

0

0

0

0

0

   bipartite

0

0

0

5

0

Rib(s):

 

 

 

 

 

   accesory

6

5

22

0

7

   Absent or less developed

0

0

0

3

8*

   wavy

2

0

0

2

3

   fused

0

0

0

4

5

Pelvic, hypoplastic to absent

0

1

0

1

1

Brachydactyly

0

0

0

3

1

Syndactyly

0

0

0

0

2

Adactyly

0

0

0

1

0

Hemimelia & small scapula

0

1

0

0

0

1Fetuses may have more than one defect

*Significantly different from control (p<0.05) by Chi-square only

Conclusions:
Subtoxic dietary levels of 0.5 and 1.0% read-across substance docusate calcium (DCS) ingested on gestational days 6 through 15 showed no adverse effects on the various maternal or fetal parameters. Toxic dietary levels of 2.0% DCS produced significant incidences of resorptions. Near toxic and toxic dietary levels of 1.5% and 2.0% DCS produced significant gross abnormalities consisting primarily of exencephaly of varying degrees with spina bifida, macroglossia, microphtalmia or anophtalmia. Since only maternally toxic does fed on gestational day 6-15 produced embryotoxic and teratogenic effects, it is concluded that no real hazard exists.
Executive summary:

Prenatal developmental toxicity was studied in rats dosed from day 6 to day 15 of gestation by dietary administration of read-across substance docusate calcium at dose levels of 0.5, 1.0, 1.5 and 2.0 % in the diet as well as by oral gavage at 0, 500 and 750 mg/kg bw.

Subtoxic dietary levels of 0.5 and 1.0% docusate calcium (DCS) ingested on gestational days 6 through 15 showed no adverse effects on the various maternal or fetal parameters. Near toxic or toxic dietary levels of 1.5 and 2.0% DCS produced significant incidences of resorptions and gross abnormalities consisting primarily of exencephaly of varying degrees with spina bifida, macroglossia, microphtalmia or anophtalmia.  Since only maternally toxic doses fed on gestational day 6-15 produced embryotoxic and teratogenic effects, it is concluded that no real hazard exists.  The dose of 1% DCS, correesponsing wiht 1060 mg/kg bw was considered to be a maternal and developmental NOAEL. This was also applied to DSS.

Endpoint:
developmental toxicity
Remarks:
rabbit as second species
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
REACH Annex IX section 8.7.2 column 2 states that a pre-natal development test shall be performed initially with one species and that a decision whether a test with a second species is needed, should be based on the outcome of the initial pre-natal development test and all available data for that substance.
Docusate sodium (CAS No. 577-11-7) has a long history of pharmaceutical and dietary uses. It is used in pharmaceutical applications such as laxative formulations, stool softener products, but also in vitamin/mineral supplements. Dietary supplements are more prone to chronic ingestion than worker exposure, therefore extensive safety data have been generated on this substance including various repeated dose, reproductive and prenatal developmental toxicity studies.
Initially, both FDA (US Food and Drug Agency) and JECFA (Joint FAO/WHO Expert Committee on Food Additives) toxicological evaluators had concerns with regard to the potential for Docusate sodium to elicit adverse reproductive or teratological effects. This possibility has been extensively examined in several studies, most notably in a multigeneration reproduction toxicity study that was conducted at the request of a Scientific Review Panel convened by FDA in 1984 with the purpose of evaluating the safety of Docusate sodium for pharmaceutical and food uses. Upon review of the full report of the three-generation study, the FDA evaluators concurred with the investigators’ conclusions that NOEL for Docusate sodium, when administered in the diets of rats for three successive generations, is 0.1% for body weight loss of parental animals and offspring, with a NOEL for reproductive parameters of 1.0%. The FDA toxicologists also concluded that reduced pup survivability observed in some litters at the 1.0% dose level was a spurious finding that was not directly related to treatment with Docusate sodium. Thus, in this study, the NOEL for Docusate sodium administration, 0.1% in the diet was based solely on a reduction in body weights for parental animals and pups.' The conclusions reached by JECFA in the appended monograph are substantively identical to FDA's.

As explained in the dossier, a supporting 2-generation generation study in rats at dietary doses of 0.5 and 1% Docusate sodium did also not demonstrate substance related effects in the parents and in the offspring. Prenatal developmental toxicity was further also tested in two studies, of which one key study with dietary administration of Docusate sodium in rats given from day 6 to 15 of gestation resulted in a maternal and developmental NOAEL of 1% (corresponding with 1074 mg/kg bw/day). The other, supporting, study with ‘Docusate calcium’ confirmed that 1% in the diet was a maternal and developmental NOAEL.

A study in the rabbit, which is the normal second species for teratogenicity testing, is not possible from a technical viewpoint. The rabbit was used in repeated dose (up to 6 month) toxicity testing but no NOAEL could be identified due to particular surfactant properties in the gastro-intestinal tract of rabbits leading to diarrhoea followed by mortality. Hence, maternal effects will occur at dosage levels far below any dosages where (secondary) developmental effects are expected. Therefore, a test with rabbits as second species is expected to provide no further scientific information.

Based on the findings from the multigeneration studies in rat and on the evaluation of the available data from the prenatal developmental toxicity studies, it is concluded that a pre-natal development test with a second species is not needed and hence this data requirement is waived according to REACH Annex IX section 8.7.2 column 2..

Sources:
• Pharmaceutical/supplementary use: Cytec Industries, Inc. - GRAS Notification for Dioctyl Sodium Sulfosuccinate, July 20, 1998
• Rabbit study: Benaglia A.E., Robinson E.J., Utley E. and Cleverdon M.A. The Chronic Toxicity of Aerosol-OT. The Journal of Industrial Hygiene and Toxicology, 1943.
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
diet
Basis for effect level:
body weight and weight gain
early or late resorptions
Dose descriptor:
NOAEC
Effect level:
1 other: %
Based on:
act. ingr.
Remarks:
diet
Sex:
male/female
Basis for effect level:
external malformations
visceral malformations
other: skeletal variations
Remarks on result:
other: secondary to high maternally toxic dose
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 074 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Klimisch 2
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

No test data were available for current substance, however read acros data were available from Docusate sodium. Justification for read across within the category of Di-ester sulphosuccinates is documented in a separate document attached in Section 13.

Teratogenicity testing

-A key study for prenatal developmental toxicity was performed in rats dosed from day 6-15 of gestation with read across substance Docusate sodium dosed at dietary dose levels of 1.0 and 2.0 % in the diet (Roell et al., 1976). The study was conducted according to OECD 414 guideline, and was considered to be reliable, adequate and relevant. Subtoxic dietary levels of 1.0% showed no adverse effects on the various maternal or fetal parameters. Toxic dietary levels of 2.0% Docusate sodium produced significant depressions in maternal weight-gains and increased incidences of resorptions (13.7%) and gross abnormalities either among litters (25.0%) or fetal populations (20.2%) as compared to controls. These abnormalities consisted primarily of exencephaly of varying degrees with, at times, spina bifida, anophthalmia and associated skeletal defects. The visceral observations confirmed the significance of the exencephalous characteristics and anophthalmia for the group given dietary levels of 2.0%. In this group, skeletal observations revealed a significant incidence of incomplete ossification to absence of the various cranial bones, a curved or open vertebral column, and a variety of defects of the vertebrae and ribs. Interpretation of the results of the present experiment, in which only maternally toxic doses induce teratogenicity, indicates no real hazard with the recommended human use of these surfactants. The concentration of 1% in the diet is considered as maternal and developmental NOAEL. This dose level corresponded with a test article intake of 1074 mg/kg body weight, as calculated in the study.

-As supporting information, prenatal developmental toxicity was also studied in rats by dietary administration of Docusate 'calcium' (DCS) at dose levels of 0.5, 1.0, 1.5 and 2.0 % in the diet as well as by oral gavage at 250, 500, 750 and 1000 mg/kg bw (Roell et all., 1976). Subtoxic dietary levels of 0.5 and 1.0% Docusate calcium ingested on gestational days 6 through 15 showed no adverse effects on the various maternal or fetal parameters. Near toxic or toxic dietary levels of 1.5 and 2.0% DCS produced significant incidences of resorptions and gross abnormalities consisting primarily of exencephaly of varying degrees with spina bifida, anophthalmia and associated skeletal defects. However, dietary levels of 2% of DCS fed to pregnant rats for 3 days (days 6-8, 8-10 or 10-12) did not produce teratogenic response. Also, DCS given to pregnant rats by oral intubation at maternally subtoxic doses (250-750 mg/kg) and a slightly toxic dose (1000 mg/kg) did not lead to malformations, however the incidence of resorptions was increased at the 2 toxic doses. Likewise doses of 500 and 750 mg/kg given by gavage from day 6-15 produced an increase in resorptions at the highest dose without a teratogenic effect. Since only maternally toxic doses fed on gestational day 6-15 produced embryotoxic and teratogenic effects, it is concluded that no real hazard exists.  

- As doscusate sodium has a long history of pharmaceutical and dietary supplement use, a 3-generation toxicity study in rat was requested by FDA and JECFA, which confirmed that docusate sodium would pose ro reproduction or teratological problems. The data were considered to adequately support safety in combination with information from prenatal developmental toxicity testing of docusate sodium and docusate calcdium, therefore a study in a second species was not considered necessary. In addition, the rabbit was not considered to be an adequate model due to its high gastro-intetinal sensitivity to surfactant properties, as demonstrated by diarrhoea followed by mortality in a repeated dose toxicity study. In conclusion, based on the evaluation of the outcome of the first species and other supporting informatoin, a second species for developmental toxicity was not considered necessary and waived according to REACH Annex IX section 8.7.2 column 2.

Conclusion

Prenatal developmental toxicity was tested by dietary administration of read across substance Docusate sodium in rats from day 6 to 15 of gestation. 1% in the diet was a maternal and developmental NOAEL corresponding to 1074 mg/kg bw, whereas at 2% in the diet visceral and skeletal anomalies were observed, which was secondary to maternal toxicity. This was confirmed in a similar study with Docusate calcium given at subtoxic and toxic dose levels, where the same could be observed.

Based on the absence of developmental findings in the teratogenicity study, no further testing is needed.


Justification for selection of Effect on developmental toxicity: via oral route:
Key study

Justification for classification or non-classification

Based on these results and according to the EC Directive (No.93/21/EEC) and CLP (No. 1272/2008 of 16 December 2008), the test substance does not have to be classified and has no obligatory labelling requirement for reproductive and developmental toxicity.