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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
reproductive toxicity, other
Remarks:
Organ toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data is from NTRL

Data source

Reference
Reference Type:
secondary source
Title:
Initial Submission: Short Term (17-Days) And Sub-Chronic (90-Days) Toxicity Studies with Tinopal CH 3669 in Rats with Cover Letter Dated 100992
Author:
Central Inst Nutrit & Food Res
Year:
1992
Bibliographic source:
National Technical Reports Library, OTS0571834,1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: as below
Principles of method if other than guideline:
Repeated Dose Toxicity Study of Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate (Tinopal CH 3669) in Rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate (Tinopal CH 3669)
- Molecular formula (if other than submission substance): C40H38N12O18S6.6Na
- Molecular weight (if other than submission substance): 1305.1422 g/mole
- Substance type: Organic
Specific details on test material used for the study:
- Name of test material (as cited in study report): Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate (Tinopal CH 3669)
- Molecular formula (if other than submission substance): C40H38N12O18S6.6Na
- Molecular weight (if other than submission substance): 1305.1422 g/mole
- Substance type: Organic

Test animals

Species:
rat
Strain:
other: Newly weaned albino
Sex:
male/female
Details on test animals and environmental conditions:
- Source: CIVO-colony
- Age at study initiation: (P) x wks; (F1) x wks: Newly weaned albino rats
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: males (40 – 61 g) and 40 females (37 - 59 g)
- Fasting period before study:
- Housing:
- Diet (e.g. ad libitum): The animals were fed on stock diet with Tinopal added at levels of 0, 0.2 and 1.0 %, ad libitum
- Water (e.g. ad libitum): Tap Water, ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 24°c

Administration / exposure

Route of administration:
oral: feed
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: stock diet
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The tent compound was thoroughly mixed into stock diet by means of a mechanical blender (type LOdige, Paderborn).

DIET PREPARATION
- Rate of preparation of diet (frequency):once a fortnight
- Mixing appropriate amounts with (Type of food): stock diet
- Storage temperature of food: At room temperature.

VEHICLE
- Justification for use and choice of vehicle (if other than water): stock diet
- Concentration in vehicle: 0, 200, 1000 and 5000 mg/Kg bw
- Amount of vehicle (if gavage):
- Lot/batch no. (if required):
- Purity:
Details on mating procedure:
not specified
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
90 days
Frequency of treatment:
Daily
Details on study schedule:
not specified
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
200 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Dose / conc.:
5 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
Total: 80
0 mg/kg/day: 10 male, 10 female
200 mg/kg/day: 10 male, 10 female
1000 mg/kg/day: 10 male, 10 female
5000 mg/kg/day: 10 male, 10 female
Control animals:
yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:
Survival, clinical sign, body weight, food consumption and water intake were examined.
Oestrous cyclicity (parental animals):
not specified
Sperm parameters (parental animals):
not specified
Litter observations:
not specified
Postmortem examinations (parental animals):
Hematology, clinical chemistry, Organ weight, gross pathology and histopathology were examined.
Postmortem examinations (offspring):
not specified
Statistics:
not specified
Reproductive indices:
not specified
Offspring viability indices:
not specified

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
When treated with 5000 mg/kg bw, all animals looked very sick in the second week of the experiment.
No abnormalities in appearance were observed in 200 and 1000 mg/kg bw treated rats.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
When treated with 5000 mg/kg bw, all animals were died at 5th week of study. Except for one female which succumbed in the 9th week.
No mortality were observed in 200 and 1000 mg/kg bw treated rats.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
When treated with 1000 and 5000 mg/kg bw, Decrease in body weight were observed in treated rats as compared to control.
When treated with 200 mg/kg bw, slight decrease in body weight but not significant as compared to control from week 6 onwards.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
When treated with 5000 mg/kg bw, Decrease in food consumption were observed in treated rats as compared to control.
When treated with 200 and 1000 mg/kg bw, slight decrease in food consumption were observed in treated rats as compared to control.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
When treated with 5000 mg/kg bw, Decrease in food efficiency were observed in treated rats as compared to control.
When treated with 1000 mg/kg bw, slight decrease in food efficiency were observed in treated rats as compared to control.
When treated with 200 mg/kg bw, decrease in food efficiency were observed in treated rats as compared to control.
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
When treated with 1000 mg/kg bw, significant increase in percentage of neutrophils and White blood cell counts were observed in treated male and female rats as compared to control.
When treated with 200 mg/kg bw, significant increase in Packed cell volume were observed in male rats but no dose-relationship was observed.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
When treated with 1000 mg/kg bw, increase in SGOT activity of females and decreased in SAP activity of males were observed as compared to control.
When treated with 200 mg/kg bw, decreased SGOT activity in males but not observed at 1000 mg/kg bw and is therefore considered incidental.
Slight decrease in G6Pase activity with increasing levels of Tinopal was observed in females only. When the enzyme activity is expressed per unit body weight the figures show a slight tendency to increase with increasing leve1s of Tinopal in males, while in females there are no distinct differences between the groups.
Very slight decrease in G6PD enzyme activity with increasing levels of Tinopal in females. Here, too, this decrease disappeared when the enzyme activity was expressed per unit body weight
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Necrosis, swelling, irregular vacuolization (Sudan III negative) and granular deterioration (eosinophylic, PAS-negative granules) of tubular epithelial cells in the renal cortex of all male and felllll1e rats at 1000 mg/kg bw.
Sloughed necrotic tubular material was often found in tubular lumens. The nuclei of viable tubular cells varied considerably in size and chromatin content. Dilated tubules lined by flattened epithelium and filled with proteinaceous casts were seen in most animals.
Moderate degree of bilateral testicular atrophy were observed in male rats at 1000 mg/kg bw.
A number of seminiferous tubules only contained Sertoli cells and occasional spermatogonia containing an irregularly vacuolated cytoplasm, Partial inhibition of spermatogenosis occurred in other tubules in which many polynucleated giant cells were seen. Varying numbers of seminiferous tubules still showed a normal development of sperm cells.
In the epididymic tubules cellular debris, scattered spermatozoa and a few giant cells were found.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
food efficiency
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
other: No effect observed

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

Dose descriptor:
other: not specified
Based on:
not specified
Sex:
not specified
Basis for effect level:
other: not specified
Remarks on result:
other: not specified

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 200 mg/kg/day for P generation when Newly weaned albino male and female rats treated with Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate (Tinopal CH 3669) orally by feed for 90 days.
Executive summary:

In a Repeated Dose Toxicity Study, Newly weaned albino male and female rats treated with Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate (Tinopal CH 3669) in the concentration of 0, 200, 1000 and 2000 mg/kg/day orally in feed for 90 days. All animals were died at 5th week of study at 5000 mg/kg bw. Except for one female which succumbed in the 9th week. No mortality was observed in 200 and 1000 mg/kg bw treated rats. All animals looked very sick in the second week of the experiment at 5000 mg/kg bw. No abnormalities in appearance were observed in 200 and 1000 mg/kg bw treated rats. Decrease in body weight were observed in treated rats at 1000 and 5000 mg/kg bw as compared to control. Slight decrease in body weight but not significant as compared to control from week 6 onwards at 200 mg/kg bw. Decrease in food consumption were observed in treated rats at 5000 mg/kg bw and slight decrease in food consumption were observed in treated rats at 200 and 1000 mg/kg bw as compared to control. Decrease in food efficiency were observed in treated rats at 5000 mg/kg bw and slight decrease in food efficiency were observed in treated rats at 1000 mg/kg bw as compared to control. Decrease in food efficiency were observed in treated rats at 200 mg/kg bw as compared to control. Similarly, significant increase in percentage of neutrophils and White blood cell counts were observed in treated male and female rats at 1000 mg/kg bw and significant increase in Packed cell volume were observed in male rats at 200 mg/kg bw but no dose-relationship was observed. Increase in SGOT activity of females and decreased in SAP activity of males were observed at 1000 mg/kg bw, decreased SGOT activity in males at 200 mg/kg bw but not observed at 1000 mg/kg bw and is therefore considered incidental. Slight decrease in G6Pase activity with increasing levels of Tinopal was observed in females only. When the enzyme activity is expressed per unit body weight the figures show a slight tendency to increase with increasing levels of Tinopal in males, while in females there are no distinct differences between the groups. Very slight decrease in G6PD enzyme activity with increasing levels of Tinopal in females. Here, too, this decrease disappeared when the enzyme activity was expressed per unit body weight. In addition, increase in relative weights of the kidney, liver and the brain and decrease in testis and ovary weight were observed at 1000 mg/kg bw as compared to control. No effect on reproductive organ weight of 200 mg/kg bw treated male and female rats were observed as compared to control. Large, pale kidneys in all male and female rats at 1000 mg/kg bw. Most of the male rats in this group had strikingly small testicles, other les ions noted at autopsy, such as unilateral hydronephrocis, eurly signs of murine pneuronia, and proteinaceous plugs in urinary bladder, are frequently encountered in the strain of used rats. They may therefore, not be related to the ingestion of the test compound. Necrosis, swelling, irregular vacuolization (Sudan III negative) and granular deterioration (eosinophylic, PAS-negative granules) of tubular epithelial cells in the renal cortex of all male and felllll1e rats at 1000 mg/kg bw. Sloughed necrotic tubular material was often found in tubular lumens. The nuclei of viable tubulur cells varied considerably in size and chromatin content. Dilated tubules lined by flattened epithelium and filled with proteinaceous casts were seen in most animals. Moderate degree of bilateral testicular atrophy were observed in male rats at 1000 mg/kg bw. A number of seminiferous tubules only contained Sertoli cells and occasional spermatogonia containing an irregularly vacuolated cytoplasm, Partial inhibition of spermatogenosis occurred in other tubules in which many polynucleated giant cells were seen. Varying numbers of seminiferous tubules still showed a normal development of sperm cells. In the epididymic tubules cellular debris, scattered spermatozoa and a few giant cells were found. Therefore, NOAEL was considered to be 200 mg/kg/day for P generation when Newly weaned albino male and female rats treated with Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate (Tinopal CH 3669) orally by feed for 90 days.