Docusate sodium

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity was tested by a key 3 -generation toxicity study and supportive 2- to 3 -generation toxicity study with Docusate sodium (DSS) performed in rats . In both studies, 1% in the diet (corresponding with 750 mg/kg bw) was a reproductive NOAEL, not resulting in impairment of fertility. Reduced body weights were observed in parentals and offspring at 1% dietary doses, however these 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%.

Link to relevant study records

Reference

Endpoint:

three-generation reproductive toxicity

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.

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

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:4DSS02 and 5DSS02
The first batch (4DSS02)was used through October 28, 1985. The second batch (5DSS02) was used from October 29, 1985 until completion of the study.


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: Stable for at least 28 days at room temperature.

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 approximately 7 wks; F1 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° (20,5 - 23,8 °C)
- 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 1,500 g 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). 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

Details on study schedule:

- F1 parental animals not mated until test diets had been fed to the animals for at least 10 weeks postweaning for all litters. Pups were allowed to nurse for 21 days before weaning. At weaning, F1 and F2 offspring were group-housed by sex by litter during the interim period elapsing between weaning of the first litter and weaning of the last litter prior to random selection of males and females for the 10-week pre-mating treatment period. All offspring were weaned directly onto their parents’ respective control or DSS diets. Breeding of the F0 generation for F1 litters was initiated on December 13, 1984 after males and females had received test material for 10 and 2 weeks, respectively. Breeding of the F1 generation for F2 litters was initiated on April 25, 1985 after test diets had been fed to the animals for at least 10 weeks postweaning for all litters. Breeding of the F2 generation for F3 litters was initiated on September 12, 1985 after the animals had received test material for at least 10 weeks postweaning for all litters.
- Selection of parents from F1 generation when pups were 21 days of age.
Four days after birth (Day 4 of lactation), the number and sex of pups, individual pup weights, and the presence of milk in the abdomen (for F2 and F3 litters) were recorded. Pups were culled from the litters at random to achieve a maximum litter size of 10, with 5 males and 5 females/litter if possible.
Pups were allowed to nurse for 21 days before weaning. At weaning, F1 and F2 offspring were group-housed by sex by litter during the interim period elapsing between weaning of the first litter and weaning of the last litter prior to random selection of males and females for the 10-week pre-mating treatment period.
- Age at mating of the mated animals in the study: minimum 13 weeks (21 days weaning and 10 weeks pre-mating)

Dose / conc.:

0 other: % nominal in diet

Dose / conc.:

0.1 other: % nominal in diet

Dose / conc.:

0.5 other: % nominal in diet

Dose / conc.:

1 other: % 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 4 days after birth (Day 4 of lactation). Pups were culled from the litters at random to achieve a maximum litter size of 10, with 5 males and 5 females/litter if possible.
Pups were allowed to nurse for 21 days before weaning. At weaning, F1 and F2 offspring were group-housed by sex by litter during the interim period elapsing between weaning of the first litter and weaning of the last litter prior to random selection of males and females for the 10-week pre-mating treatment period.

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.

Oestrous cyclicity (parental animals):

Not examined.

Sperm parameters (parental animals):

No sperm investigation performed.

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 10 pups/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

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: All surviving animals after litters had been weaned

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

HISTOPATHOLOGY / ORGAN WEIGHTS
Microscopic examination of the collected tissues was not required.

Postmortem examinations (offspring):

SACRIFICE
- The F1 and F2 offspring not selected as parental animals and 20 weanlings/sex/group of the F3 litters were sacrificed at 5 days of age (Day 4 of lactation).

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. 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

Remarks:

Generat: maternal/paternal toxicity

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

Key result

Dose descriptor:

NOAEC

Remarks:

reproduction/offspring

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

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

Remarks:

maternal/paternal toxicity

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

Dose descriptor:

NOAEC

Remarks:

reproduction/offspring

Effect level:

1 other: %

Based on:

act. ingr.

Remarks:

diet

Sex:

male/female

Basis for effect level:

other: no effects up to highest concentration

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:

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:

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.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

750 mg/kg bw/day

Species:

rat

Quality of whole database:

reliable

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

A key study 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 similar 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 parents and offspring of all generations. 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 -to 3-generation generation study in rats, dietary doses given were 0.5 and 1% (Cytec, Levinskas & Shaffer 1970). 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 (F3a). 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.

Effects on developmental toxicity

Description of key information

Prenatal developmental toxicity was tested by a key dietary administration with Docusate sodium (DSS) in rats from day 6 to 15 of gestation and a supporting dietary administration study of Dosucsate calcium (DCS) . In both studies, 1% in the diet (corresponding with 1074 mg/kg bw) was a maternal and developmental NOAEL, whereas at higher concentrations (2% for DSS and 1.5 and 2% for DCS) in the diet developmental toxicity was observed, which was considered secondary to maternal toxicity. The higher dosages were exceeding the limit dose of 1000 mg/kg bw and resulted in excessive maternal toxicity.

Testing in a second species was not considered ethically acceptable as Docusate salts have been used for a long time as pharmaceutical agent and ingredient, and etensive data were available in humans. No increased risk of malformations was concluded in epidemiological studies from more than 800 patients (pregnant women) which were available for Docusate sodium (or other salts) as pharmaceutical, mainly used during the first trimester of pregnancy. In the Adverse Drug Reaction database from EMA (up to May 2021), a total of 933 Adverse Drug reactions (ADRs) were reported, however for pregnancy, puerperium and perinatal conditions, only 26 ADRs were reported of which 5 were considered serious. The serious cases were most likely not due to Docusate sodium, but to other comedication.

Various publications pointed out the same conclusion, and the outcome was considered to be reliable and extensive.

o further testing is considered needed based on these data.

Link to relevant study records

Reference

Reference 1

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.

In its Final Deciision Letter of 14 November 2019 (Decision number: CCH-D-2114489574-33-01/F), ECHA remarked an information gap and pointed to the necessity to provide information for this endpoint. The rabbit might not be a suitable species for testing (due to the surfactant properties in the GI tract of rabbits leading to diarrhoea followed by mortality); ECHA acknowledged this concern and agreed the study should be conducted in an appropriate second species (e.g. mouse). Since the substance to be tested is a solid, ECHA concluded that testing should be performed by the oral route.

As a follow-up, a thorough search was performed in Public literature and (Adverse Drug Reaction) databases, which resulted in extensive pharmaceutical use of Docusate sodium or other salts, as active ingredient under multiple brand names up to 500 mg daily by oral application, and as inactive ingredient up to a maximum daily exposure of 50 mg for oral use. Various epidemiological data are available for Docusate sodium use by pregnant women, of which the majority were exposed in the first trimester of pregnancy (a total of 821 pregnant women were exposed during pregnancy, for which no increased risk of malformations was reported). In the Adverse Drug Reaction database from EMA (2021), a total of 933 Adverse Drug Reactions (ADRs) were reported, however for pregnancy, puerperium and perinatal conditions, only 26 ADRs were reported of which 5 were considered serious. The serious cases were most likely not due to Docusate sodium, but to other comedication. Various publications pointed out the same conclusion, and the outcome was considered to be reliable and extensive.

Based on these findings, further testing in a second species was not considered ethically acceptable by the Consortium as Docusate salts have already been used for a long time as pharmaceutical agent and ingredient, and etensive data were available in humans, including high numbers of pregnant women. No increased risk of malformations was concluded in epidemiological studies from more than 800 patients (pregnant women) which were available for Docusate sodium (or other salts) as pharmaceutical, mainly used during the first trimester of pregnancy. The limited ADRs in the target population of pregnant women in the more recent database Adverse Drug Reaction database from EMA (up to May 2021) confirmed the literature findings.

More information is provided in the attached literature study under the Endpoint summariy of Section 7.10, and under the separate endpoint records of 7.10. Exposure related information in humans.

Species:

rabbit

Reference 2

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.

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

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No. of test material: DSS was obtained from American Cyanamid Company as Control Lot #7535

FORM AS APPLIED IN THE TEST (if different from that of starting material): DSS was prepared as a 40% solution in (USP) corn oil.

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:

dosing from gestational days 6 through 15

Frequency of treatment:

continuous in the diet

Duration of test:

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

Dose / conc.:

1 other: %

Remarks:

nominal in diet; Exp. II-A.

Dose / conc.:

2 other: %

Remarks:

nominal in diet; Exp. II-B.

Dose / conc.:

1 074 mg/kg bw/day (actual dose received)

Remarks:

1.0%: calculated compound consumed

Dose / conc.:

1 988 mg/kg bw/day (actual dose received)

Remarks:

2.0%: calculated compound consumed

No. of animals per sex per dose:

43 pregnant female rats in control group
22 pregnant female rats in dose 1.0%
20 pregnant female rats in dose 2.0%

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Adult rats can tolerate, at least, 1% of DSS in their diets (equivalent to about 600 - 700 mg/kg/day) for 1 year without adverse effects on any of the various parameters tested including histopathology (Fitzhugh and Nelson, 1948). The purpose of this investigation was to evaluate the possible teratogenic effects of DSS when given orally to pregnant rats during periods of gestational development, and examination of offspring for abnormalities .

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: all per litter
- 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.

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

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):

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 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:

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.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 074 mg/kg bw/day

Species:

rat

Quality of whole database:

reliable

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available (further information necessary)

Additional information

A key study for prenatal developmental toxicity was performed in rats dosed from day 6-15 of gestation with docusate sodium (DSS) 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 external abnormalities either among litters (25.0%) or fetuses (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 variations included 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 dose induced developmental toxicity, 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 corresponds 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 variations. Since only maternally toxic does fed on gestational day 6-15 produced developmental effects, it is concluded that no real hazard exists.   The present experiment, in which only maternally toxic dose induced developmental toxicity, 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 corresponds with a test article intake of 1060 mg/kg body weight, as calculated in the study.

Based on ECHA request for prenatal development evaluation in a second species; human data of Docusate (sodium) used during pregnancy were investigated. According to the ECHA Guidance on Information Requirements (Chapter R.7a: Endpoint specific guidance Version 6.0 - July 2017), human data on reproductive and developmental toxicity can be used, either based on epidemiological data/studies, case reports and clinical data. As Docusate (sodium) has been used extensively as pharmaceutical agent or ingredient, testing in a second species (e.g. mouse as proposed by ECHA as an alternative to rabbit) was considered not ethically feasible. Therefore a waiver for a second species was justified based on available data that have been entered under Section 7.10 Health surveillance data, epidemiological data and exposure related observations in humans. A summary is provided below, whereas the more detailed endpoints are available in Sections 7.10 (1/2/5).

Various epidemiological data are available in literature for Docusate sodium use by pregnant women, of which a smaller group was exposed anytime (N = 116) during pregnancy, whereas most were exposed in the first trimester of pregnancy. However, two publications used the same data, one including pregnancies with life births only and the other all pregnancies. A corrected total of 821 pregnant women (705 during first trimester) exposed to Docusate (sodium) was derived. In total, the first trimester was therefore studied in a group of >700, for which no increased risk of malformations was reported (incidence = 0.2 - 3.9%). An overview of literature studies and No. of pregnant women is provided below. The studies summarised were considered to be reliable (Klimisch 2).

-       N = 116 anytime during pregnancy (Prospective): No increased risk of malformations (Heinonen et al., 1977)

-       N = 473 during first trimester (Surveillance): No increased risk of malformations (1/473 = 2%) (Jick et al., 1981) #, *

-       N = 319 during first trimester (Surveillance): No increased risk of malformations (3/319 = 0.9%) (Aselton et al., 1985) #, **

-       N = 232 during first trimester (Surveillance): no increased risk of malformations (9/232 = 3.9%) (Briggs et al., 2011) §

Total = 1140 (1024 during first trimester) Corrected = 821 (705 during first trimester)

# Studied Based on GHC (Group Health Cooperative) of 6,837 pregnant women

* Drugs Prescrobed During the First Trimester of Pregnancy to at Least 200 of 6,837 Pregnant Women

** Drugs Prescribed During the First Trimester of Pregnancy to at Least 200 of 6,509 Women Having Live Births Studied

§ In a surveillance study of Michigan Medicaid recipients involving 229,101 completed pregnancies conducted between 1985 and 1992, 232 newborns had been exposed to a docusate salt during the 1st trimester (F. Rosa, personal communication, FDA, 1993). Nine (3.9%) major birth defects were observed (nine expected), including one cardiovascular defect (two expected) and one polydactyly (one expected). No anomalies were observed in four other categories of defects (oral clefts, spina bifida, limb reduction defects, and hypospadias) for which specific data were available. These data do not support an association between the drug and congenital defects.

Docusate (sodium) still seems amongst the most commonly used over-the-counter medication components (Drugs.com; Shafe et al., 2011). Doses in pregnant women vary from 50 to 500 mg/day orally in one to four divided doses or 0.12 g rectally as active docusate sodium in a 10 g enema gel. Most frequently used as docusate salts, sodium docusate and calcium docusate, although other forms are available (Rungsiprakarn et al., 2015). Docusate has not been formally assigned to a pregnancy category by the FDA (Drugs.com).

Docusate sodium is available under multiple brand names. In order to prevent and treat chronic constipation or as an adjunct in abdominal radiological procedures, Docusate sodium should be taken up by adults (p.o.) up to 500 mg daily in divided doses. Treatment should be commenced with large doses, which should be decreased as the condition of the patient improves. According to the FDA inactive ingredient list (2021), Docusate sodium is listed up to a maximum daily exposure of 50 mg for oral use. In the Adverse Drug Reaction database from EMA (2021), a total of 933 ADRs were reported (May/2021). However, for pregnancy, puerperium and perinatal conditions, only 26 ADRs were reported of which 5 were considered serious. The serious cases were most likely not due to Docusate sodium, but to other comedication.

Conclusion

No increased risk of malformations was concluded in epideùiological studies from more than 800 patients (pregnant women) which were available for Docusate sodium (or other salts) as pharmaceutical, mainly used during the first trimester of pregnancy.

Further exploration of literature and databases (e.g. FDA Inactive Ingredient List, EMA Adverse Drug Reaction database) confirmed that Docusate sodium is available as active ingredient under multiple brand names up to 500 mg daily by oral application, and as inactive ingredient up to a maximum daily exposure of 50 mg for oral use. In the Adverse Drug Reaction database from EMA (2021), a total of 933 Adverse Drug Reactions (ADRs) were reported, however for pregnancy, puerperium and perinatal conditions, only 26 ADRs were reported of which 5 were considered serious. The serious cases were most likely not due to Docusate sodium, but to other comedication. No further testing is considered needed based on these data.

Justification for classification or non-classification

As there were only secondary effects due to paternal/maternal toxicity, classification for reproductive and developmental toxicity is not indicated.