Registration Dossier

Administrative data

Description of key information

Acute oral toxicity with test substance containing >= 90% act. ingr. was tested by gavage in 3 studies, showing LD50 between 580 and 1400 mg/kg for male and female rats in the key study; the test material was therefore considered harmful if swallowed. Acute dermal toxicity testing in rats did not reveal relevant

changes and resulted in an LD50 > 2000 mg/kg bw. Inhalation toxicity testing was waived based upon the fact that acute inhalation exposure as such is very unlikely for sulfosuccinates due to their substance properties.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1986-1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to GLP and valid testing guidance, however limited data on test substance identification were provided. Nevertheless, the study is reliable, relevant and adequate for classification.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
No. of animals
Qualifier:
equivalent or similar to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Version / remarks:
79/831/EWG, Annex V, Part B
Principles of method if other than guideline:
2-5 animals/sex/dose; Orientation test.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Borchen
- Age at study initiation: young
- Weight at study initiation: fasted mean weight 165-201 g
- Fasting period before study: ca. 16 hours before and 3 hours after application
- Housing: 2-5 animals per Makrolon 3 cage, soft wood granulates(ARWI-Center, Essen) as bedding
- Diet (e.g. ad libitum): Altromin-Haltungsdiät 1324 (Fa. Altromin GmbH, 4937 Lage) ad libitum
- Water (e.g. ad libitum): Public supply, ad libitum
- Acclimation period: 7 or 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 20-25°C
- Humidity (%): ca. 45-60%
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: 25.111986-03.02.1987
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 10%, 7.0% and 2.9% (w/v)

MAXIMUM DOSE VOLUME APPLIED: 20 mL/ kg
Doses:
2000mg/kg; 1400 mg/kg; 580 mg/kg
No. of animals per sex per dose:
2 high dose
5 mid dose and low dose
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Symptoms and mortality several times at the day of application, thereafter twice daily. Body weight 1 day before application;before application (fasted) on the day of application; 48 hours, 7 and 14 days after application.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology

Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
> 580 - < 1 400 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 580 - < 1 400 mg/kg bw
Based on:
test mat.
Mortality:
Dose 2000 mg/kg: 2/2 male and 2/2 female animals died 24 hours after application.
Dose 1400 mg/kg: 4/5 male animals died 24 hours after application and 5/5 female animals died 6-24 hours after application.
Dose 580 mg/kg: 0/5 male animals died 14 days after application and 1/5 female died 24hours-14 days after application.
Clinical signs:
Dose 2000 mg/kg:
Piloerection and decreased activity was seen in all male and female animals (4/4) after ca. 1 h.
Piloerection, decreased activity and diarrhea was seen in 2/2 males after ca. 2 h and in 2/2 females after ca. 2-5 h.
Piloerection, decreased activity, diarrhea, squatting attitude and decreased body temperature was seen in 2/2 males after ca. 5 h.
Dose 1400 mg/kg:
5/5 male animals showed moderate piloerection, slightly decreased activity, curved gait, squatting attitude and decreased body temperature after ca.2-3 hours. 5/5 males showed strong piloerection, moderate to marked decreased activity, otherwise unchanged symptoms after ca.3-6 hours.
5/5 female animals showed moderate piloerection, slightly decreased activity, curved gait, squatting attitude and decreased body temperature after ca. 2-3 hours. 4/5 females showed strong piloerection, decreased activity, otherwise unchanged symptoms after ca. 3-6 hours.
Dose 580 mg/kg:
5/5 male animals showed moderate piloerection after ca. 2-4,5 hours. 2/5 males showed diarrhea after ca. 2 hours. 5/5 female animals showed moderate piloerection, moderate curved gait and slightly decreased activity after ca. 2-4,5 hours. 1/5 female showed diarrhea after ca. 2 hours.
Body weight:
See Under Tables below.
Gross pathology:
Dose 2000 mg/kg:
2/2 male animals had a gastro-intestinal tract high-grade filled with liquid, mucosa partially reddish discolored and low-grade emphysema. 2/2 female animals had a bloodily nose and muzzle, otherwise the same symptoms as the males.
Dose 1400 mg/kg:
4/5 males showed strongly reddish discolored medulla, emphysema of the lungs, accumulation of liquid in the thick and small intestine, 1/5 showed no pathological signs. 5/5 female animals showed strongly reddish discolored medulla, emphysema of the lungs, accumulation of liquid in the thick and small intestine, and in addition 1/5 female showed low-grade bleedings in the fundus area.
Dose 580 mg/kg:
The male animals had no special findings. 3/5 females had no special findings. 1/5 female had moderate hydrometra, 1/5 female showed a strongly reddish discolored medulla, emphysema of the lungs and accumulation of liquid in the thick and small intestine.

Males: mean body weights (g) Females: mean body weights (g)

Dose (mg/kg 2000 1400 580 2000 1400 580

-1 day 212 207 209 178 179 181

Start experiment 201 196 197 165 169 171

+ 2 days - 186* 212 - - 175

+ 7 days - 217* 237 - - 178

+ 14 days - 250* 267 - - 186

Body weight gain - 43 58 - - 5

*= value of one survivor

Interpretation of results:
other: acute harmful
Remarks:
Criteria used for interpretation of results: Off. J. Europ. Comm., L 257, 1983, p.18
Conclusions:
The oral LD50 of the test item containing ≥ 90% active ingredient was <1400 mg/kg >580 mg/kg for male rats
and <1400 mg/kg >580 mg/kg for female rats.
Executive summary:

The acute oral toxicity of test item containing ≥90% act. ingr. was tested by oral gavage in young Wistar rats at dose levels of 580, 1400 and 2000 mg/kg bw. The test compound was administered by single gavage in aqua dest. as solvent and an application volume of 20 mL/kg bw to fasted animals. Two (low and mid dose) or five rats (high dose) were used per sex and dose. Clinical observations and gross macroscopic observations were observed at all dose levels . The LD50was <1400 mg/kg and >580 mg/kg for male rats and <1400 mg/kg and >580 mg/kg for female rats. According to “Off. J. Europ. Commun., L 257, 1983, p. 18”, the test item can be classified as acute harmful.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
580 mg/kg bw
Quality of whole database:
High quality (Klimish 2)

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to GLP and valid testing guidance, therefore it is considered relevant, adequate and reliable for classification.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: CD / Crl: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: Approx. 8 weeks
- Weight at study initiation: Males (at dosing): 227–254 g; Females (at dosing): 201-226 g
- Fasting period before study: Approx. 16 hours before administration; only tap water was then available ad libitum
- Housing: During the 141-day observation period the animals were kept singly in MAKROLON cages (type III plus). Granulated textured wood (Granulat A2, J. Brandenburg, 49424 Goldenstedt, Germany) was used as bedding material for the cages. The cages were changed and cleaned twice a week.
- Diet (e.g. ad libitum): Commercial diet, ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany)
- Water (e.g. ad libitum): Drinking water in bottles was offered ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C (maximum range)
- Humidity (%): 55% ± 15% (maximum range)
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 7,2012 To: September 11, 2012
Type of coverage:
occlusive
Vehicle:
other: Aqua ad iniectabilia
Details on dermal exposure:
TEST SITE
- Area of exposure: intact dorsal skin on the animals back between the fore and hind extremities ; an area of at least 5 cm x 6 cm
- % coverage: approx. 1/10 of body surface
- Type of wrap if used: The test item was held in contact with the skin with 8 layers of gauze. The gauze was covered with a plastic sheet and secured with adhesive plaster strips on the application site for 24 hours.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): No

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10mL/kg bw (a correction factor of 1.05 was used)
- Concentration (if solution): 2.1 g/ g water
- Constant volume or concentration used: constant concentration, different volume (3.1 g/kg bw of the test item preparation)
- For solids, paste formed:


Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Observations were performed before and immediately, 5, 15, 60 and 60 minutes, as well as 3, 6 and 24 hours after administration. All animals were observed for a period of 14 days. During the follow-up period (two weeks), changes of skin and fur, eyes and mucous membranes, respiratory and circulatory function, autonomic and central nervous system and somatomotor activity as well as behaviour pattern, were observed at least once a day until all symptoms subsided, thereafter each working day. Attention was also paid to possible tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma.
Observations on deaths were made at least once daily to minimize loss of animals during the study.
Individual body weights were recorded before administration of the test item and thereafter in weekly intervals up to the end of the study.
- Necropsy of survivors performed: yes
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
act. ingr.
Mortality:
None of the animals died prematurely.
Clinical signs:
A very slight erythema (barely perceptible) on the application site was observed in all 5 of 5 male and 5 of 5 female animals on test days 2 and 3.
Body weight:
All animals gained the expected body weight throughout the whole experimental period.
Gross pathology:
No macroscopic findings were observed at necropsy.
Interpretation of results:
not classified
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The test item (>95% purity) requires no labelling (as LD50 > 2000 mg/kg bw).
Also, according to the EC Regulation 1272/2008 and subsequent regulations, the test material is not classified for acute dermal toxicity.
Executive summary:

In this experiment, the test item (>95% purity) was examined for acute toxicity after a single dermal application to rats at one dose level of 2000 mg act. ingr./kg bw. The test item was applied once for 24 hours on the shaved intact dorsal skin of rats (5 cm x 6 cm, approx. 1/10 of body surface). This treatment was followed by an observation period of 2 weeks. Under the present test conditions, a single dermal administration of 2000 mg/kg bw to rats revealed no signs of toxicity and no deaths. A very slight erythema (barely perceptible) on the application site was observed in all 5 of 5 male and 5 of 5 female animals on test days 2 and 3. All animals gained the expected body weight throughout the whole experimental period. No macroscopic findings were observed at necropsy.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
High quality (Klimisch 1)

Additional information

Acute oral toxicity

- A key study for acute oral toxicity for the registered substance was available with test item containing >= 90% active ingredient dosed by gavage in Wistar rats at 480, 1400 and 2000 mg/kg bw (Potokar, 1987). Five (for low and mid dose) and two rats (for high dose) were used per sex. Clinical observations and gross macroscopic observations were observed at all dose levels. The LD50 was between 580 and 1400 mg/kg for male and female rats, and therefore the test item was considered harmfull.

- Two supporting studies for registered substance for acute oral toxicity by gavage were available for other test batches:

1. Test item containing >= 90% act. ingr. was tested by oral gavage in male and female Wistar rats (Potokar, 1986).

The male LD50 was > 2000mg/kg bw; the female male LD50 was ca. 1400 mg/kg bw.

2. Test item containing >= 90% act. ingr. was tested by oral gavage in male Wistar rats (Gloxhuber, 1969). The test item was administered by single gavage in aqua dest. as solvent and an application volume of 20 mL/kg bw to fasted animals at doses of 1580, 1990, 2510 and 3160 mg/kg bw. Ten male rats were used per sex and dose. The LD50 was 2400 mg/kg bw.

In conclusion, the test item is considered of low toxic potential based on the most detrimental study, indicating an LD50 between 580 and 1400 mg/kg bw. For the LD50 value, 580 mg/kg was used as worst case value.

 

Acute dermal toxicity

A key study for acute dermal toxicity for registered substance was conducted with test substance containing >95% active ingredient (Haferkorn, 2013). One dose level of 2000 mg/kg bw was employed. The test item was applied once for 24 hours on the shaved intact dorsal skin of rats (5 cm x 6 cm, approx. 1/10 of body surface). This treatment was followed by an observation period of 2 weeks. There were no signs of toxicity and no deaths. A very slight erythema (barely perceptible) on the application site was observed in all 5 of 5 male and 5 of 5 female animals on test days 2 and 3. All animals gained the expected body weight throughout the whole experimental period. No macroscopic findings were observed at necropsy.

In conclusion, as LD50 exceeds 2000 mg/kg bw and only very slight erythema was observed, there is no acute dermal toxicity hazard.

 

Acute inhalation toxicity

Inhalation is very unlikely due to large particle size, low vapour pressure and high hydrophilic properties of the substance.  Based on these and other physicochemical properties, the inhalation route is not appropriate; the oral and dermal route of administration are therefore applied as first and second relevant routes (ECHA R7a Guidance p 342). Additional inhalation testing would therefore neither lead to a better risk assessment, nor improve the safety of applications. On the basis of the argumentation summarized above an acute inhalation toxicity study is waived.

 

Conclusion

Acute oral toxicity with test substance containing >= 90% act. ingr. was tested by gavage in 3 studies, showing LD50 between 580 and 1400 mg/kg for male and female rats in the key study; the test material was therefore considered harmful if swallowed. Acute dermal toxicity testing in rats did not reveal relevant changes and resulted in an LD50 > 2000 mg/kg bw. Inhalation toxicity testing was waived based upon the fact that acute inhalation exposure as such is very unlikely for sulfosuccinates due to their substance properties.

Justification for selection of acute toxicity – oral endpoint  Key study 

Justification for selection of acute toxicity – dermal endpoint  Key study

Justification for classification or non-classification

The test substance is classified according to CLP regulation (No. 1272/2008 of 16 December 2008) as Category 4 for acute oral toxicity with signal word 'WARNING', H302.

Based on these results and according to CLP (No. 1272/2008 of 16 December 2008), the test substance does not have to be classified and has no obligatory labelling requirement for dermal toxicity.