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Administrative data

Description of key information

NOAEL 90 days rat ≥ 300 mg/Kg bw/day

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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test procedures cannot be subsumed under a testing guideline, nevertheless they are well documented and scientifically acceptable.
Principles of method if other than guideline:
A subchronic test of 13 weeks was performed in rats. Groups of 15 males and 15 females were administered by oral gavage with 30, 100 and 300 mg/kg bw/day of test substance in water emulsion (Cremophor).
GLP compliance:
no
Remarks:
pre GLP
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Weight at study initiation: males 114 and females 120 g.
- Housing: in groups of 5 in Macrolon cages (type 3).
- Diet: ad libitum, Altromin-R-Food.
- Water: ad libitum, tap water.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C.
- Photoperiod: from the 7.00 a.m. to 7.00 p.m.
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
water emulsion (Cremophor).
Duration of treatment / exposure:
91 days.
Frequency of treatment:
Daily.
Remarks:
Doses / Concentrations:
30, 100 and 300 mg/kg
Basis:
actual ingested
No. of animals per sex per dose:
15 males and 15 females x dose x treated group.
30 males and 30 females in the control group.
Control animals:
yes, concurrent vehicle
Details on study design:
Control group dosed with 5.0 ml/kg of vehicle.
Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS
The animals were inspected daily.

BODY WEIGHT
The body weight was measured weekly.

HAEMATOLOGY
The following blood tests were performed in 5 rats pre dose after 13 weeks: hemoglobin as cyanmethemoglobin, counting the erythrocytes and leukocytes, platelet count, hematocrit determination, calculation of Färbe-coefficient, calculation of the corpuscular volume, assessment of complete blood count based on smears.

CLINICAL CHEMISTRY
In 5 rats of all groups, the following determinations were carried out after 13 weeks: activity of serum glutamate pyruvate Transeminase (GPT), serum glutamic-oxaloacetic transaminase, serum sorbitol dehydrogenase, glutamate dehydrogenase. In addition, the bilirubin concentrations were determined in serum.
In 5 rats of all groups, the concentration of urea and creatinine were determined after 13 weeks in the serum.

URINALYSIS
The urine were examined for albumin, sugar and blood; bile pigments: Ehrlich's reagent, Schlesinger's reagent; microscopic sediment analysis; specific gravity.
Sacrifice and pathology:
After 13 weeks, the rats were anesthetized and killed by exsanguination.
The internal organs (thyroid, heart, Lungs, liver, spleen, kidneys, adrenals, testes and Ovarian) were macroscopically evaluated and weighed.
Clinical signs:
no effects observed
Description (incidence and severity):
no difference from the control group
Mortality:
no mortality observed
Description (incidence):
no difference from the control group
Body weight and weight changes:
no effects observed
Description (incidence and severity):
compatible with the control group
Haematological findings:
no effects observed
Description (incidence and severity):
all values ​​found were within the physiological range
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Description (incidence and severity):
no pathological findings
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
absolute and relative organ weights did not differ fron control
Gross pathological findings:
no effects observed
Description (incidence and severity):
no changes were recorded
Details on results:
CLINICAL SIGNS AND MORTALITY
During the three month test period the rats of the treated groups did not differ from those of the control group.

BODY WEIGHT AND WEIGHT GAIN
The body weight increase in the treated groups was compatible with that recorded in the control group.

HAEMATOLOGY
All values ​​found were within the physiological range.

CLINICAL CHEMISTRY
No indication of liver damage. For the kidney functionality evaluation, the values ​​found were all in the normal range.

URINALYSIS
Also in the urine no pathological findings were observed.

ORGAN WEIGHTS
The average values ​​of the absolute and relative organ weights of the groups of rats treated with the drug did not differ significantly from those of the control group.

GROSS PATHOLOGY
The internal organ macroscopically analysis showed no changes.
Dose descriptor:
NOAEL
Effect level:
>= 300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Critical effects observed:
not specified
Conclusions:
NOAEL ≥ 300 mg/kg bw/day (nominal).
Executive summary:

Method

A subchronic test of 13 weeks was performed in rats. Groups of 15 males and 15 females were administered by oral gavage with 30, 100 and 300 mg/kg bw/day of test substance in water emulsion (Cremophor).

Results

During the three month test period the rats of the treated groups did not differ from those of the control group. The body weight increase in the treated groups was compatible with that recorded in the control group.

No treatment related findings were recorded in the hematological, clinical chemestry analysis and urinalysis. The internal organ macroscopically analysis showed no changes.

Conclusion

NOAEL ≥ 300 mg/kg bw/day (nominal).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subchronic
Species:
rat

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

The substance under registration (CAS 67786-25-8) belongs to the category of Stilbene Fluorescent Whitening Agents.

This substance and all other members of this category do not show acute toxic effects after oral, inhalation, and dermal administration. They are neither irritant to skin nor eyes, nor genotoxic in-vitro and in-vivo, nor sensitizing. Six over fourteen registered substances were tested for subchronic toxicity and four of them were tested for chronic toxicity up to two years with no relevant toxic effects (further details are given in the Category Justification Report, Section 13).

 

In a preliminary screening/dose range finding on the substance under registration CAS 67786-25-8 30 male and 30 female rats were administered orally for five days. On the first day of the experiment they were administered with 200 mg/kg; in the following four days the dose was increased by a factor of 1.5 (300, 450, 675 and 1015 doses were administered, respectively). Observation period was extended one week after the last dosing. The results indicate no cumulative effect of the compound. (NOEL ≥ 1015 mg/kg (nominal)) (Kimmerle and Solmecke, 1972).

A subchronic test of 13 weeks was then performed in rats. Groups of 15 males and 15 females were administered by oral gavage with 30, 100 and 300 mg/kg bw/day of test substance in water emulsion (Cremophor). During the three month test period the rats of the treated groups did not differ from those of the control group. The body weight increase in the treated groups was compatible with that recorded in the control group. No treatment related findings were recorded in the haematological, clinical chemistry analysis and urinalysis. The internal organ macroscopically analysis showed no changes. The NOAEL was established as greater than 300 mg/kg bw/day (nominal) (Kimmerle and Solmecke, 1972).

 

Reliable data on repeated dose toxicity subacute and chronic after oral exposure of rats are available for CAS 16470-24-9 and CAS 4404-43, the tetrasulphonated sodium salt and disulphonic acid form, respectively.

 

The substance CAS 16470-24-9 can be considered as the best representative within the category, since it has the same sulphonation degree than the substance under registration CAS 67786-25-8, a very high structural similarity (Tanimoto distance > 0.9), very similar molecular weight (1165

Vs 1121 da of CAS 67786-25-8) and a very high water solubility (650 g/l Vs 1000 g/l of CAS 67786-25-8).

Based on the metabolic pathway proposed by the OECD Toolbox can be also expected that the same conclusions about kinetic and excretion can be drawn and applied to CAS 67786-25-8 as well as for CAS 16470-24-9: the two substances differ in the fact that the substitution on the triazino moiety is a dihydroxyethylamine for CAS 16470-24-9 and a bis(2-hydroxypropyl) for CAS 67786-25-8. No breakdown metabolisation products are proposed for CAS 67786-25-8, therefore based on the higher water solubility and the metabolic pattern a rapid and complete excretion is expected.

The reason for the proposed lowering in metabolic activity can be found in the fact that the hydroxyl group is in the registering case a secondary one and more sterically hindred. Secondary alcohols are excreted primarily as the glucuronic acid conjugates (Williams, 1959[1]; Lington & Bevan, 1994 [2]).

Toxicological results for many other endpoint were demonstrated as similar (acute aquatic toxicity, acute oral, irritation, sensitisation, genotoxicity). In conclusion, Read Across between CAS 67786-25-8 and CAS 16470-24-9 is justified.

 

In a chronic toxicity study (equivalent to OECD 453, Bomhard et al., 1978) the test substance CAS 16470-24-9 was administered to 50 Wistar rats/sex/dose in diet at dose levels of 0, 100, 1000, 10000 ppm (10000 ppm = 709 mg/kg bw/day for females and 521 mg/kg bw/day for males) for 24 months. There were no compound related effects in mortality, clinical signs, body weight, food consumption, haematology, clinical chemistry, urinalysis, organ weights, or gross and histological pathology. Therefore, the NOAEL was determined for females at 709 mg/kg bw/day and for males at 521 mg/kg bw/day. This chronic study in the rat is acceptable and satisfies the guideline requirements for a chronic oral study OECD 453 in rats.

The same chronic study (Bomhard E. and Löser E., 1978) was also performed on the acid form of the disulphonated derivative (CAS 4404-43-7). Appearance, behaviour, feed intake, body weights and mortality were not influenced in male and female animals of doses up to and including 10000 ppm. The animals in the dose groups of 10000 ppm did not show during the entire experimental period any treatment-related symptoms. The growth of the rats was not affected until the dose of 10000 ppm. The haematological investigations performed during and at the end of the test showed no dose of injuries. The clinical chemical analysis, sections and histopathological examinations revealed no evidence for treatment-related damage to the liver. Urinalysis, urea and creatinine concentrations in serum as well as macroscopic and histopathological organ findings did not indicate any influence. NOAEL: 779 mg/kg bw/day (actual dose received) (female) NOAEL: 542 mg/kg bw/day (actual dose received) (male).

 

As representative value for the endpoint the NOAEL 90 days rat of 300 mg/kg bw/day obtained on the substance under registration has been considered.

REFERENCE

[1]Williams, R. T. (1959) Detoxification Mechanisms. The Metabolism and Detoxification of Drugs, Toxic Substances and Other Organic Compounds, 2nd Ed., London: Chapman & Hall, pp. 4, 114–126.

[2]Lington, A. W. & Bevan, C. (1994) Alcohols. In: Clayton, G. D. & Clayton, F. E., eds, Patty’s Industrial Hygiene and Toxicology, 4th Ed., New York: John Wiley & Sons, Vol. IID, pp. 2585–2760.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
A 90 day study was performed on the substance according to international accepted guideline.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.9 Specific target organ toxicity - repeated exposure section, substances are classified as specific target organ toxicants following repeated exposure by the use of expert judgement, on the basis of the weight of all evidence available, including the use of recommended guidance values which take into account the duration of exposure and the dose/concentration which produced the effect(s), and are placed in one of two categories, depending upon the nature and severity of the effect(s) observed.
Classification in Category 2 is applicable, when significant toxic effects observed in a 90-day repeated-dose study conducted in experimental animals are seen to occur within the guidance value ranges as:
- oral (rat): 10 < C ≤ 100 mg/kg bw/day

The No Observed Adverse Effect Level was established at 300 mg/kg bw/day, on the basis of the results from a subchronic study of 13 weeks.

 

In conclusion, the available experimental data are adequate for classification and labelling purpose and the substance is not classified for repeated dose toxicity according to the CLP Regulation (EC 1272/2008).