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Toxicological information

Repeated dose toxicity: oral

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Administrative data

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 and the validity of the study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1969
Report Date:
1969

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
(only one dose per test substance)
GLP compliance:
no
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
- Name of test material (as cited in study report): Aerosol MA, sodium dihexylsulfosuccinate
- Physical state: Not provided. It is deduced that source material was a viscous paste.
- Analytical purity: 78-80%
- Impurities (identity and concentrations): See confidential details
- Composition of test material, percentage of components: See confidential details
- Purity test date: Not provided
- Lot/batch No.: Lot#W-90109, SPS##7997, Notebook#S8484, Page 54
- Expiration date of the lot/batch: Not provided
- Stability under test conditions: Not provided
- Storage condition of test material: Not provided

Test animals

Species:
rat
Strain:
other: Charles River strain albino
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River Breeding Laboratories, North Wilmington, Mass.
- Age at study initiation: Not provided
- Weight at study initiation: 100 g (female rats), 121 g (male rats) on average
- 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.
- Water: No data provided
- Acclimation period: Not provided

ENVIRONMENTAL CONDITIONS
Not provided


Administration / exposure

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 20g/rat/day
(Derelanko M.J., 2008, The Toxicologist's Pocket Handbook, Informa).
1% in the diet = 10000 mg/kg diet corresponds with 10 mg/g diet
20 g feed/rat (250g bw)/day = 80 g feed/kg bw/day = 0.8 g active ingredient/kg bw/day = 800 mg/kg bw/day.
A higher feed intake is possible, e.g. 1000 mg/kg at higher body weight and feed intake, but from a conservative viewpoint 750 mg/kg bw is taken


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


Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
1.0%
Basis:
nominal in diet
No. of animals per sex per dose:
For Aerosol MA: 20 male+20 female at 1.0% dietary level + 20 male and 20 female as control
Control animals:
yes, concurrent no treatment
Details on study design:
Experimental Animals
The animals employed in the study were Charles River strain ( Charles River Breeding Laboratories, North Wilmington, Mass.) albino rats. Two hundred and eighty rats (140 males and 140 females) were selected for the experiment, ear-punched with the animal number assigned and housed individually in standard wire-bottomed steel rat cages. Each cage bore a color-coded card identifying the animal with respect to project number, test material assignment, individual animal number and sex.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: Yes

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

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined : Yes
and mean daily diet consumption calculated as g food/kg body weight/day: No
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: 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 checked in table [No.IV and V] were examined.
Hematocrit Value
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(fasted serum glucose concentration)
- How many animals: 5 rats of each sex (=10) and 10 control
- Parameters checked in table [NoVI and VII] were examined.
Blood Urea Nitrogen (BUN) Concentration
Serum Alkaline Phosphatase (SAP) Activity
Serum Glutamic-Pyruvic Transaminase (SGPT) Activity
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 checked in table [No.VIII] were examined.
Glucose Concentration
Albumin Concentration
Microscopic Elements Examination
pH
Specific Gravity



NEUROBEHAVIOURAL EXAMINATION: Yes , but in general, not specific
- Time schedule for examinations: abnormal reactions and death were recorded daily during the investigation
- Dose groups that were examined: control and 1.0% dose
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No

Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
ORGAN WEIGHTS: Yes
Other examinations:
No
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.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
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
Details on results:
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

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).

Effect levels

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Dose descriptor:
NOAEL
Effect level:
1 other: % in the diet
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No toxicological findings (only a slight significant absolute liver weight decrease in males)
Dose descriptor:
NOAEL
Effect level:
ca. 750 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No toxicological findings (only a slight significant absolute liver weight decrease in males)

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The comparisons 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.
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.
There were no significant differences between the tissues of test and control rats observed upon histopathological examination.
Executive summary:

Six groups of 40 albino rats (20 male, 20 female Charles River Strain) plus 1 control group (20 male, 20 female) were fed with 1% of various test items mixed into the diet. The various test items were category members of the Sulfosuccinates Diester Group including Butanedioic acid, sulfo-, 1,4-bis (1,3-dimethylbutyl)ester, sodium salt. 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. No significant differences in clinical blood chemistry studies and absolute organ weights have been detected. 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 category members at 1% in the diet (10000 ppm equivalent to 750 mg/kg body weight/day on average basis) for 90 days in rats did not result in any relevant changes in the subchronic toxicity study. The NOAEL was therefore considered to be ca. 750 mg/kg bw/day.

The IBT Report, supplemented by the Intox report and the Validation Report of October 15, 1983, may be considered a valid study and the data and conclusions relied upon.