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

Endpoint summary

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.

Repeated dose toxicity was tested in various species, including rats, dogs, rabbits and monkeys.  The NOAEL of 750 mg/kg bw/day obtained in the key study in rats was confirmed to be consistent with  data from docusate sodium and category members in supporting studies in rats; other data from other species were of limited reliability and relevance and therefore not taken into account.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1969

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to internationally accepted test guidelines and is considered relevant, adequate and reliable. There were some deviations from the study guidelines, however these did not affect the conclusions reached and the validity of the study.

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 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

only one dose per test substance

GLP compliance:

no

Limit test:

yes

Species:

rat

Strain:

other: Charles River strain albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Charles River Breeding Laboratories, North Wilmington, Mass.
- Weight at study initiation: 101-122 g
- Housing: individually in standard wire-bottomed steel rat cages
- Diet : standard rat ration blended with the appropriate amount of test material in a Hobart Mixer
Fresh diets were prepared each week. Each rat was offered an amount of diet sufficient for one
week ‘ad libitum’ feeding. However, checks were made periodically to ensure that the food jars were
not empty

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: 1.0% in the feed
Taking into account a mean body weight of 250 g and a mean food consumption of 25g/rat/day:
10000 mg/kg diet corresponds with 1000 mg/kg bw/day on average basis:
25 g feed/rat (250g bw)/day = 100 g feed/kg bw/day = 1g active ingredient/kg bw/day = 1000 mg/kg bw/day.

DIET PREPARATION
- Rate of preparation of diet (frequency): Fresh diets were prepared each week
- Mixing appropriate amounts with (Type of food): standard rat ration


VEHICLE: No

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

90 days

Frequency of treatment:

continuously

Remarks:

Doses / Concentrations:
10.000 ppm (1% in the diet)
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
750 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

20 males & 20 females

Control animals:

yes

Positive control:

no

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: no

BODY WEIGHT: Yes
- Time schedule for examinations: biweekly (day 0, 15, 30, 45, 60, 75 and 90).

FOOD CONSUMPTION AND COMPOUND INTAKE: yes
- Food consumption for each animal determined : yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: no

FOOD EFFICIENCY: no

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: yes
- Time schedule for collection of blood: after 84 days
- Anesthetic used for blood collection No data
- Animals fasted:yes (fasted serum glucose concentration)
- How many animals: 5 rats of each sex (=10) and 10 control
- Parameters: Hematocrit, Erythrocyte Count, Hemoglobin Concentration, Total Leukocyte Count, Differential Leukocyte Count

CLINICAL CHEMISTRY: yes
- Time schedule for collection of blood: after 84 days
- Animals fasted: yes
- How many animals: 5 rats of each sex (=10) and 10 control
- Parameters: Blood Urea Nitrogen (BUN), Serum Alkaline Phosphatase (SAP), Serum Glutamic-Pyruvic Transaminase (SGPT), Fasted Serum Glucose Concentration

URINALYSIS: yes
- Time schedule for collection of urine: after 84 days
- Metabolism cages used for collection of urine: No data
- Animals fasted: yes
- Parameters: Glucose Concentration, Albumin Concentration, Microscopic Elements Examination, pH, Specific Gravity

NEUROBEHAVIOURAL EXAMINATION: no

Sacrifice and pathology:

ORGAN WEIGHTS AND ORGAN TO BODY WEIGHT RATIO'S: yes
- organs: liver, kidney

GROSS AND HISTOPATHOLOGY
Following 90 days of feeding, all surviving rats were sacrificed by carbon dioxide asphyxation and autopsied. Animals which died during the study were examined grossly unless examination was precluded by post-mortem autolysis. At the time of gross examination a complete set of organs and other tissues was removed from each rat and preserved in formalin solution. Also at autopsy the weight of the liver and kidneys of ten rats of each sex in every group was determined and recorded.
Microscopic examination of tissues taken from five rats of each sex in every group was conducted. The following tissues, stained with Hematoxylin-Eosin, were included: esophagus, stomach (cardia, fundus and pylorus), small intestine (duodenum, jejunum and ileum), cecum, colon, liver, kidneys, spleen, pancreas, urinary bladder, pituitary gland, adrenal gland, testes, seminal vesicle, ovary, bone marrow, thyroid gland, parathyroid gland, salivary gland, prostate gland, heart, aorta, lung, lymph node (cervical and mesenteric), skeletal muscle, peripheral nerve, bone (femur), spinal cord, uterus, trachea, eye, optic nerve and brain (cerebrum, cerebellum and pons).

Statistics:

Statistical analyses were conducted upon the absolute organ weights and their corresponding ratios to the weight of the body. An Analysis of Variance was conducted first and any significant effects disclosed by that treatment were further studied by “t” –tests.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

See Table 2

Food consumption and compound intake (if feeding study):

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Dose descriptor:

NOAEL

Effect level:

750 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

clinical signs

Critical effects observed:

not specified

Table 2. Body Weight Data - Summary of Mean Values

		Body Weight (grams)							Total Weight Gain (grams/rat)
Group	Sex	Day 0	Day 15	Day 30	Day 45	Day 60	Day 75	Day 90
Control	M	122	178	254	340	403	435	466	344
	F	101	154	188	212	239	273	280	179
OT	M	121	180	253	338	398	424	428	307
	F	101	152	180	209	241	256	262	161

 

Conclusions:

The comparison of final body weights and total weight gains revealed no statistically significant differences between test and control animals.
No outstanding differences in food consumption were noted between test rats and control rats.
No deaths or abnormal behavioral reactions were noted among any of the animals employed in the study.
No outstanding differences between test and control rats were noted with respect to any of the blood parameters studied.
(With the exception of an elevation of both SGPT and SAP values among males fed Aerosol TR,) all data obtained from test rats were not different than those from control animals.
No significant differences between the urine of test rats and control rats were observed.
No outstanding differences between test and control rats were noted at the time of gross pathological examination.
( The only statistically significant difference noted was smaller absolute liver weights among male rats fed 1.0% Aerosol IB.)

Executive summary:

No repeat dose toxicity 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.

A group of 40 albino rats (20 male, 20 female Charles River Strain) plus 1 control group (20 male, 20 female) were fed with 1% Docusate sodium mixed into the diet. After 84 days 5 hematological values, 4 blood chemical values, 5 urinalysis values were measured for all animals. 40 tissues have been examined pathologically at the conclusion of the 90-days test period. Organ to body weight and organ to brain weight ratios were calculated. There were no significant differences in clinical blood chemistry studies and absolute organ weights. Body weights organ to body weight ratios, hematologic studies and urinalysis were not different between test and control animals. No deaths or abnormal behavioral reactions occurred; no gross pathological findings were noted.

Administration of 1% Docusate sodium in the diet (10000 ppm equivalent to 750 mg/kg body weight/day) for 90 days in rats did not result in any relevant changes in the subchronic toxicity study. The NOAEL was therefore considered to be 750 mg/kg bw/day. The IBT Report, supplemented by the Intox report and the Validation Audit Report of October 15, 1983, may be considered a valid study and the data and conclusions relied upon.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

750 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Reliable

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

A key study for oral subchronic toxicity was performed in rats with Docusate sodium (Cytec, Plank 1969a); the study was conducted according to internationally accepted test guidelines, and was considered to be reliable, adequate and relevant. The validity of the study was supported by additional audits on the raw data and histopathological evaluation. Although deficiencies were detected compared to current standards, the study was concluded to be valid and reliable. A group of 40 albino rats (20 males and 20 females) was fed with 1% test substance mixed into the diet, compared to a control group. There were no significant differences in body weights, haematology, clinical blood chemistry, urinalysis and organ weights. No deaths or abnormal behavioral reactions occurred; no gross pathological findings were noted. Administration of 1% docusate sodium in the diet (10.000 ppm equivalent to approximately 750 mg/kg body weight/day) for 90 days in rats did not result in any relevant changes in the subchronic toxicity study. The NOAEL was therefore established at 750 mg/kg bw/day.

In that same study, various category members/structural analogues were tested as supporting information at the same doses: Sodium Diamylsulfosuccinate, Sodium Dicyclohexylsulfosuccinate, Diisobutylsulfosuccinate, Sodium Dihexylsulfosuccinate, Sodium Di- (tridecyl) sulfosuccinate (Cytec, Plank 1969b). Five groups of 40 albino rats were fed with 1% test substance mixed into the diet for the same parameters as listed above, compared to a control group. With the exception of an evaluation of both SGPT (serum glutamic pyruvic transaminase) and SAP (serum alkaline phosphatase) values among males fed CAS 2673-22-5 and smaller absolute liver weights among males fed CAS 127-39-9, no significant differences in clinical blood chemistry studies and absolute organ weights have been detected. Administration of 1% in the diet (10.000 ppm equivalent to approximately 750 mg/kg body weight/day) for 90 days in rats did not result in any relevant changes in the subchronic toxicity study. The NOAEL was 750 mg/kg bw/day for the various category members.

In a supporting study, groups of 10 (5 male & 5 female) Wistar rats were treated for 6 months at concentrations of 0.25, 0.5, 0.75, 1.0 and 1.25 g/kg diet, corresponding to doses of190, 370, 550, 750 and 870mg/kg bw/day. Occasional spells of diarrhea occurred in some animals, particularly at the higher doses. Neither the total red cell, total white cell, nor the differential counts of rats was affected by the continued administration . The dose level of 750 mg/kg was confirmed as NOAEL (Literature, Benaglia et al. 1943).

Other studies were also available from literature in various species (Literature, Benaglia et al. 1943 and Case et al., 1977) in dogs, rabbits and Rhesus monkeys. The other species were considered to be less appropriate due to the gastrointestinal tensio-active local irritation by which systemic effects could not be fully evaluated. The NOAEL of 750 mg/kg bw/day in rats was selected as main endpoint, as this was confirmed in various studies and with different category substances.

In summary, repeated dose toxicity was tested in various species, including rats, dogs, rabbits and monkeys. The NOAEL of 750 mg/kg bw/day obtained in the key study in rats was confirmed to be relevant and consistent.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

Key study

Repeated dose toxicity: via oral route - systemic effects (target organ) digestive: colon

Justification for classification or non-classification

As the NOAEL for the read-across substance sodium docusate is higher than treshold levels given in the classification guidelines, there is no indication for the need to classify the registered substance docusate calcium.