Calcium 1,4-bis(2-ethylhexyl) bis(2-sulphosuccinate)

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Link to relevant study records

Reference

Reference 1

Endpoint:

three-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1984-10-04 To 1985-12-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study has been extensively conducted, but has not all parameters as requested in the guideline.

Justification for type of information:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Principles of method if other than guideline:

No sperm investigation, estrus cycle, organ weights, histopathology.

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl: CD(SD) BR purchased as weanlings

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts (Portage, Michigan facility)
- Age at study initiation: (P=F0) x approximately 7 wks; (F1) x in utero
- Fasting period before study: No
- Housing: individually in suspended, stainless steel, screen bottom cages, held on racks, with
deotized animal cage boards (DACB, Shepherd Specialty Papers, Inc., Kalamazoo, Michigan)lining
the urine- and feces-collecting pans.
During mating periods, double-sized stainless steel cages were used to house one male and one
female rat. Cage boards were changed at least twice weekly and animals were transferred to clean
cages at least biweekly except during the mating period. Pregnant rats (during the last week of
gestation) and rats with young, as well as females that did not exhibit evidence of positive mating,
were individually housed in plastic cages fitted with water bottles filled with tap water. A bedding
material consisting of heat-treated hardwood chips (Beta-Chip, Northeastern Products Corporation,
Warrensburg, New York) covered the bottom of the cages, and was changed at least once
weekly. Variations from these procedures are documented in the raw data.
- Diet : ad libitum from glass containers that limit spillage and contamination and allow easy
inspection of the amount and condition of the feed. The basal diet for this study was ground Purina
Certified Rodent Chow #5002. A record of all lot numbers is included in the raw data. This diet has
and the minimum or maximum concentrations of these materials are specified.
- Water : partially demineralized by reverse osmosis and sterilized by ultraviolet light, ad libitum
from an automatic watering system (Systems Engineering, Palo Alto, California), except during the
last week of gestation and during lactation when tap water was provided in water bottles to
animals in plastic cages. Hazleton Laboratories America, Inc. (HLA) analyzed the tap water and
water from the automatic system quarterly for total dissolved solids, conductivity, heavy metals,
organophosphates, chlorinated hydrocarbons, microbiological content and selected elements.
Results of these analyses were within normal limits and are on file at HLA.
- Acclimation period: at least 4 weeks

ENVIRONMENTAL CONDITIONS
- Temperature : 72°F±3°
- Humidity (%):50%±20%
- Air changes (per hr): at least 10 changes an hour, filtered 100% outside air

Route of administration:

oral: feed

Vehicle:

acetone

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Approximately 1.500g of the weighed rodent chow was
transferred to a Hobart N50. DSS was added to a measured amount of acetone and the admixed with
the basal diet in the Hobart N50 for 5 minutes. This premix was then added to the rodent chow in
the Hobart H600T mixer and mixed for 15 minutes to achieve the proper concentration in the feed.
Each dose level for the study was prepared independently, first the low dose and then progressively
higher doses.

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Certified Purina Rodent Chow® #5002
- Storage temperature of food: mixed feed was stored refrigerated in air-tight polyethylene buckets.

VEHICLE
- Justification for use and choice of vehicle (if other than water): acetone

Details on mating procedure:

- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1
- Length of cohabitation: maximum 21 days
- Vaginal smears were taken daily to demonstrate mating.
- After 21 days of unsuccessful pairing females were placed in a plastic cage
- Further matings after two unsuccessful attempts: not applicable
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of gestation

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyses of the prepared diets were done prospectively (i.e., before feeding to the animals) using the
analytical method in Appendix B. This method is based on the hydrolysis of DSS to 2-ethyl- hexanol
which is then quantitated by gas-liquid chromatography. Analyses were performed weekly for the
first 4 weeks, weekly during the 2-week premating phase for F0 females, twice during breeding for
F1 litters, twice during F1 gestation, twice during F1 lactation, and monthly thereafter.
Because the analytical method for DSS is based on measuring a possible degradation product (2-ethyl
hexanol) , it was necessary to determine if measurable quantities of 2-ethyl hexanol were present in
nonhydrolyzed diet samples. To do this, several sets of diets were analyzed for DSS and 2-ethyl
hexanol after 8 days at ambient conditions and/or after being stored frozen for 1 to 14 days.

Duration of treatment / exposure:

F0 generation animals received the test material at the appropriate dose levels in the diet continuously, from approximately 7 weeks of age (October 4, 1984 for males; November 29, 1984 for females), throughout mating, gestation, and lactation and until they were necropsied on March 4, 5, and 6, 1985.
F1 and F2 generation animals were exposed to the test material in utero, while nursing, continuously from when they weaned to test diets , throughout mating, gestation, and lactation, and until they were necropsied on July 22, 23, and 24, 1985 (F1 animals) and on December 26 and 30, 1985 (F2
animals.

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
0%, 0.1%, 0.5%, and 1.0%
Basis:
nominal in diet

No. of animals per sex per dose:

Dosing (F0, F1 and F2) : 30/sex/group (0%,0.1%,0.5%,1.0%)

Control animals:

yes

Details on study design:

- Dose/Route selection rationale: The oral route by diet was based upon human oral exposure.
- Rationale for animal assignment: the selection of F1 pups from each litter was started after each litter had been exposed for 5 days to the respective diets.

Positive control:

No

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: males: weekly
females: weekly during pre-mating phases; on Days 0, 7, 14,and 20 of
gestation; and on Days 0, 7, 14, and 21 of lactation

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined (g ): Yes
- Compound intake, (mg/kg/day): Yes


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice of all adults (F0, F1 and F2): after litters have been weaned
- Organs examined:colon, duodenum, epididymides, (gross lesions), ileum, jejunum, kidneys, liver,
mammary gland with skin, ovaries, prostate, seminal vesicles, stomach, testes, uterus, vagina.

OTHER:
- One male and one female F1 pup were selected at random from each litter previously weaned onto their respective diets and designated to be used for a possible recovery phase.

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 10pups/litter (5/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
[number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, other:]

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals (after litters had been weaned)
- Maternal animals: (after litters had been weaned)

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

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at 4 days of age.

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

Statistics:

Differences between treated groups and the control group were considered statistically significant at p≤0.05 and are indicated with an asterisk(*).
Data for adult animals were analyzed by sex. Body weight; body weight gain; food consumption; reproduction indices; precoital interval; length of gestation; pup viability; body weights, body weight gains, and sex ratios; and litter size (alive and dead by sex) were analyzed separately using one-way nalyzes of variance (ANOVA) with transformation when necessary (log10, square, square root, reciprocal, arcsine rank) to attain variance homogeneity. If the ANOVA was significant, Dunnett’s test was used to compare each treated group with the control. When no transformation could establish variance homogeneity (as indicated by Levene’s test at p<0.01), all data were also examined by nonparametric techniques. These statistics included the Kruskal-Wallis H-test analysis of variance and, if this test was significant, the Nemenyi (F0 and F1 generations) or the Nemeneyi-Kruskal-Wallis (F2 generation) test for multiple comparisons or the Wilcoxon-Mann-Whitney two-sample rank test. Reproduction indices and the total number of live and dead pups were analyzed by the Cochran-Armitage test for trend and Fisher-Irwin exact test for heterogeneity.

Reproductive indices:

Mating = number of females mated( =number of females with vaginal sperm or plug, or
that gave birth to a litter)x100/ number of females placed with males

Female fertility = number of females pregnant(=number of females that gave birth to a litter )x100/
number of females mated

Male fertility = number of males shown to be fertile(=a female giving birth to a litter)x100 /
number of males placed with females

Gestation = number of live litters born x100/ number of pregnant females

Offspring viability indices:

Viability * = (number of pups surviving to Day 4)x100/ number of pups born alive

Weaning+ = (number of pups surviving to Day 21)x100/ number of pups alive at Day 4 postculling

Sex ratio = ( number of male pups )x100/ total number of pups

* Also reported as “Survivability Days 0-4”
+ Also reported as “Survivability Days 4 (postculling)-21”

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.

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.

Food consumption and compound intake (if feeding study):

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.

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Test substance intake: Test article intake was calculated.

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

(number of females pregnant (that gave birth to a litter) / number of females mated)x100

Reproductive function: sperm measures:

no effects observed

Description (incidence and severity):

(number of males shown to be fertile (defined by a female giving birth to a litter)/number of males placed with females)x100

Reproductive performance:

no effects observed

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:

clinical signs

Remarks on result:

other: Generation: maternal/paternal (migrated information)

Dose descriptor:

NOAEC

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

in the diet

Sex:

male/female

Basis for effect level:

clinical signs

Remarks on result:

other: Generation: offspring (migrated information)

Remarks on result:

not measured/tested

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Initial pup weights (i.e. on lactation day 0) were significantly lower than those of controls only for the high-dose group during the third generation. However, lower pup weight gains in het mid- and high-dose groups resulted in significantly lower pup we

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

not specified

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)

Remarks on result:

not measured/tested

Remarks on result:

not measured/tested

Reproductive effects observed:

not specified

Conclusions:

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:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

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% 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. There were no other effects on parental or reprocutive 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.

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.

Reference 2

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

Study period:

1970

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study has been extensively conducted, but has not all parameters as requested in the guideline.

Justification for type of information:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Limit test:

no

Species:

rat

Strain:

other: Elias strain, albino

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet: each week
- Mixing appropriate amounts with: ground PURINA® LABORATORY CHOW® (Ralston
Purina Co., St. Louis, Missouri) on a weight basis.

VEHICLE
- Justification for use and choice of vehicle (if other than water): No vehicle

Details on mating procedure:

- Impregnation procedure: - If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: maximum 21 days
-
- Further matings after two unsuccessful attempts: not applicable
- Proof of pregnancy: vaginal plug / sperm in vaginal smear]referred to as day 0 of gestation

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Young albino rats were given 0.5% and 1% 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.

Remarks:

Doses / Concentrations:
0.5% and 1.0%
Basis:
nominal in diet

No. of animals per sex per dose:

16 pairs (1male=1female) for mating, for each dietary level (0.5% and 1.0%) and for the controlgroup
litter size was reduced to 10 pups

Control animals:

yes

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: males: weekly
females: weekly during pre-mating phases; on Days 0, 7, 14,and 20 of
gestation; and on Days 0, 7, 14, and 21 of lactation

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined (g ): Yes
- Compound intake, (mg/kg/day): Yes


POST-MORTEM EXAMINATIONS: Yes
F1: intact animals observed for gross defects and discarded without autopsy
F2: intact animals observed for gross defects and discarded without autopsy
F3: Pups at 21 days of age of the first litter were autopsied. From each litter the 2 smallest males and females received special attention. One of two was cleared with potassium hydroxide and the bones wer stained with alizarin for sleketal examination. The other was processed for histology of all major organs. The pups of the second litter were observed fro gross defects and discarded without autopsy.
All animals (F0-F3) which died before weanin were exmained (without auopsy).

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not examined

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Dose descriptor:

NOAEC

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

in the diet

Sex:

male/female

Remarks on result:

other: Generation: all (migrated information)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

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 fox 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 jnvolved ;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.

Reproductive effects observed:

not specified

Conclusions:

It is concluded that feeding of 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:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, AEROSOL OT (docusate sodium; CAS: 577 -11 -7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Young albino rats were given 0.5% and 1% 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

Dose descriptor:

NOAEL

750 mg/kg bw/day

Additional information

A key study for reproductive toxicity was performed in a 3-generation toxicity study with docusate sodium at dietary dose levels of 0.1, 0.5 and 1.0% in the diet (Cytec, 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 generation study in rats, dietary doses given were 0.5 and 1% (Cytec, 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 he 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.

 

Short description of key information:

Multigeneration studies were performed to assess 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.

Effects on developmental toxicity

Description of key information

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Prenatal developmental toxicity was tested by dietary administration of 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.  

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

only 2 doses tested. Allthough not all details on the study design were provided, the study was performed to the highest standards at the time of conduct. In the current study, a concurrent test article, dioctyl calcium sulfosuccinate (DCS) was also tested at 0.5, 1, 1.5 and 2% in the diet.

Justification for type of information:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: about 2 months
- Housing: individually in hanging wire mesh cages
- Diet (e.g. ad libitum): Wayne Lab Meal ad libitum
- Water (e.g. ad libitum): ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24±2°C
- Humidity (%): 50±5%
- Photoperiod (hrs dark / hrs light):12/12

Route of administration:

oral: feed

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: 40% solution in corn oil

DIET PREPARATION
- Mixing appropriate amounts with (Type of food): 1.0% and 2.0% admixed in Wayne Lab Meal

VEHICLE
- Concentration in vehicle: 40% solution in corn oil

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Not provided

Duration of treatment / exposure:

gestational days 6 through 15: dosing

Duration of test:

gestational days 6 through 15: dosing
gestational day 21: killing of the mothers and removing fetuses by cesarean section

Remarks:

Doses / Concentrations:
1.0 and 2.0%
Basis:
nominal in diet

No. of animals per sex per dose:

22 female rats in dose 1.0%
20 female rats in dose 2.0%

Control animals:

yes, concurrent vehicle

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes (clinical condition and signs of illness)
- Time schedule: each day

BODY WEIGHT: Yes (weight gain)
- Time schedule for examinations: day 6-15; day 15-21

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
- Compound intake calculated as time-weighted averages from the consumption and body weight
gain data: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #21
- Organs examined: Ovaries and uterine content

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of resorptions: Yes

Fetal examinations:

- External examinations: Yes:
- Soft tissue examinations: No
- Skeletal examinations: Yes: [one half of the total number of fetuses]
- Head examinations: No
-Visceral examinations: Yes (one-half of the total number of fetuses were fixed in Bouin’s fluid for a detailed examination of visceral anomalies, using the slicing method of Wilson)

Statistics:

Maternal body-weight gains, maternal food consumptions and fetal weights were analyzed by Dunnett’s two-sided, multiple comparison test. Frequencies of resorptions and fetal abnormalities among litters were analyzed by the Mann-Whitney U test or the Chi-square test (with Yate’s correction), as appropriate.

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: 2.0% in the diet

Dose descriptor:

NOAEC

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

in the diet

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEC

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

in the diet

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Details on embryotoxic / teratogenic effects:
See Table 1.

Remarks on result:

other: See Table 1

Abnormalities:

not specified

Developmental effects observed:

not specified

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
Avarage, 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 malformations^1: 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 observations^1:	Group (I-A)	(II-A)	(II-B)
Total number examined	388^a	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	165^a	98	91
Defects^1:   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	167^a	97	98
Defects^1:   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% 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:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Prenatal developmental toxicity was studied in rats dosed from day 6 to day 15 of gestation by dietary administration of 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.

Reference 2

Endpoint:

developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Study period:

1976

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

In the current study, a concurrent test article, dioctyl calcium sulfosuccinate (DCS) or docusate calcium was tested at 0.5, 1, 1.5 and 2% in the diet. Allthough not all details on the study design were provided, the study was performed to the highest standards at the time of conduct.

Justification for type of information:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: about 2 months
- Housing: individually in hanging wire mesh cages
- Diet (e.g. ad libitum): Wayne Lab Meal ad libitum
- Water (e.g. ad libitum): ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24±2°C
- Humidity (%): 50±5%
- Photoperiod (hrs dark / hrs light):12/12

Route of administration:

other: via the diet and by oral gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: 40% solution in corn oil

DIET PREPARATION
- Mixing appropriate amounts with (Type of food): 1.0% and 2.0% admixed in Wayne Lab Meal

VEHICLE
- Diet: 40% solution in corn oil
- Gavage: in 48% corn oil

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Not provided

Duration of treatment / exposure:

gestational days 6 through 15: dosing
Different dosing schedules (see No. of animals/sex/dose)

Frequency of treatment:

daily

Duration of test:

gestational days 6 through 15: dosing
gestational day 21: killing of the mothers and removing fetuses by cesarean section

Remarks:

Doses / Concentrations:
0.5, 1.0, 1.5 and 2.0%
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
250, 500, 750, 1000 mg/kg bw/day
Basis:
actual ingested
gavage

No. of animals per sex per dose:

See further details on study design

Control animals:

yes, concurrent vehicle

Details on study design:

47 female control rats

Exp. I-A: day 6-15
29 female rats in dose 0.5%
38 female rats in dose 1.0%
32 female rats in dose 1.5%
38 female rats in dose 2.0%

Exp. III-A:
21 female rats in dose 2.0% - day 6-8
21 female rats in dose 2.0% - day 8-10
22 female rats in dose 1.0% - day 10-12

Exp. IV-A:
7 female rats dosed at 250 mg/kg - day 8-10
7 female rats dosed at 500 mg/kg - day 8-10
7 female rats dosed at 750 mg/kg - day 8-10
8 female rats dosed at 1000 mg/kg - day 8-10
7 female rats dosed at 1500 mg/kg - day 8-10
7 female rats dosed at 1500 mg/kg - day 10-12

Exp. V-A:
21 female rats dosed at 1000 mg/kg - day 6-8
21 female rats dosed at 1000 mg/kg - day 8-10
21 female rats dosed at 1000 mg/kg - day 10-12

Exp. VI-A:6-15
20 female rats dosed 0 mg/kg - day 6-15
21 female rats dosed at 500 mg/kg - day 6-15
23 female rats dosed at 7500 mg/kg - day 6-15

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes (clinical condition and signs of illness)
- Time schedule: each day

BODY WEIGHT: Yes (weight gain)
- Time schedule for examinations: day 6-15; day 15-21

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
- Compound intake calculated as time-weighted averages from the consumption and body weight
gain data: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #21
- Organs examined: Ovaries and uterine content

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of resorptions: Yes

Fetal examinations:

- External examinations: Yes:
- Soft tissue examinations: No
- Skeletal examinations: Yes: [one half of the total number of fetuses]
- Head examinations: No
-Visceral examinations: Yes (one-half of the total number of fetuses were fixed in Bouin’s fluid for a detailed examination of visceral anomalies, using the slicing method of Wilson)

Statistics:

Maternal body-weight gains, maternal food consumptions and fetal weights were analyzed by Dunnett’s two-sided, multiple comparison test. Frequencies of resorptions and fetal abnormalities among litters were analyzed by the Mann-Whitney U test or the Chi-square test (with Yate’s correction), as appropriate.

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: 2.0% in the diet

Dose descriptor:

NOAEC

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

in the diet

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEC

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

in the diet

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Details on embryotoxic / teratogenic effects:
See Table 1.

Abnormalities:

not specified

Developmental effects observed:

not specified

Table 1. Maternal and fetal results of pregnant rats given various amounts if 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 malformations^1: 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 observations^1:
Total number examined	388^a	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
Defects^1:   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	167^a	120	143	113	141
Defects^1:   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% 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 0.5 and 2.0% DCS produced significant incidences of resorptions and gross abnormalities consisting primarily of exencephaly of varying degress with spina bifida, anophtalmia and associated skletal defects. However, dietary levels of 2% of DCS fed to pregnant rats for 3 days (days 6-8, 8-10, 10-12) did not produce teratogenic response. Also, DCS given to pregnant rats by oral intubation at maternally subtoxic doses (250-750 mg/kg), a slightly toxic dose (1000 mg/kg ), 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.

Executive summary:

No data are available for the registered substance, however data are available for the corresponding sodium salt of the substance, docusate sodium (CAS 577-11-7). These data are used for read-across to the registered substance calcium docusate (CAS 128 -49 -4). The presence of either of the counterions sodium or calcium is not considered to have an impact on the toxicity. It is therefore considered justified to use the data of the sodium salt for read across to the calcium salt of docusate.

Prenatal developmental toxicity was studied in rats dosed from day 6 to day 15 of gestation by dietary administration of 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 250, 500, 750 and 1000 mg/kg bw. 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, anophtalmia and associated skeletal defects. However, dietary levels of 2% of DCS fed to pregnant rats for 3 days (days 6-8, 8-10, 10-12) did not produce teratogenic response. Also, DCS given to pregnant rats by oral intubation at maternally subtoxic doses (250-750 mg/kg), a slightly toxic dose (1000 mg/kg ), 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.  

Effect on developmental toxicity: via oral route

Dose descriptor:

NOAEL

1 074 mg/kg bw/day

Additional information

A key study for prenatal developmental toxicity was performed in rats dosed from day 6-15 of gestation with docusate sodium dosed at dietary dose levels of 1.0 and 2.0 % in the diet (Cytec, 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 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. 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 does fed on gestational day 6-15 produced embryotoxic and teratogenic effects, it is concluded that no real hazard exists.  

Justification for classification or non-classification

As there were only secondary effects due to paternal/maternal toxicity with the read-across analogue docusate sodium, classification for reproductive toxicity of the registered substance (docusate calcium) is not indicated.