Registration Dossier

Administrative data

Description of key information

A key study for oral acute toxicity was performed in mice, demonstrating LD50 of 11000 mg/kg bw. A key study for dermal acute toxicity was available from read across substance Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts, showing LD50 >2000 mg/kg bw. Inhalation toxicity testing was waived based upon the fact that acute inhalation exposure as such is very unlikely for sulphosuccinates due to their substance properties and the risk management measures that are already implemented.

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:
1967
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
7 days instead of 14 days observation
Principles of method if other than guideline:
Single oral application to mice and calculation of 7 day LD50
Study performance before implementation of corresponding international guidelines, however study performance is comparable to recent guidelines to great extent.
GLP compliance:
no
Remarks:
study performance before implementation of GLP
Test type:
standard acute method
Limit test:
no
Species:
mouse
Strain:
NMRI
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Institut für industrielle und biologische Forschung, GmbH, Köln
- Age at study initiation: Not provided
- Weight at study initiation: Median weight of 23.7 g
- Fasting period before study: Approximately 17-20 hours
- Housing: Prior to the application in Plastibox cages (42x28x17 cm3) in groups of 35 animals on vermiculite. After application in Makrolon cages on wire roasting over vermiculite.
- Diet (e.g. ad libitum): Altromin R, pellet diameter 8-9mm, supplier: Altromin GmbH
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: Not provided

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 21°C
- Humidity (%): ca. 70%
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): Not provided


Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.0464, 0.215, 0.464, 1.00, 1.00 and 1.00 (mL/mL) at the dose levels of 1.00, 4.64, 10.0, 21.5, 31.6 and 46.4 mL/kg bw, respectively


MAXIMUM DOSE VOLUME APPLIED: 46.4 mL/kg bw or rather 1.10 mL/mouse

Doses:
1.00 mL/kg bw; 4.64 mL/kg bw ; 10.0 mL/kg bw ; 21.5 mL/kg bw ; 31.6 mL/kg bw and 46.4 mL/kg bw
No. of animals per sex per dose:
10 for the 1.00; 4.64; and 10.0 mL/kg dose
20 for the 21.5 and 46.4 mL/kg dose
16 for the 31.6 mL/kg dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 days
- Frequency of observations and weighing: The mice were observed after application about 8 hours long at short intervals. Further assessments were performed several times daily until 7 days after application.
- Necropsy of survivors performed: yes
- Other examinations performed: vitality, pain reflex, cyanosis, breathing, corneal reflex, lid wide, exophtalmus, sound sensitivity, shaky walking, abdominal position, side position, running motion, clonic-tonic convulsions, ophistotonos, defecation; necropsy of deceased animals
Statistics:
The quantitative evaluation of the lethality data was performed by graphical determination of the LD50 7 days after application, in the probability grid with logarithmic shared X-axis, when using confidence limits.
Sex:
male
Dose descriptor:
LD50
Effect level:
28.7 mL/kg bw
Based on:
test mat.
Remarks on result:
other: LD50 as reported in study report referring to the test substance as delivered by the sponsor
Key result
Sex:
male
Dose descriptor:
LD50
Effect level:
ca. 11 000 mg/kg bw
Based on:
other: solid content
Remarks on result:
other: Recalculated LD50 referring to solid content of 39% (according to producer information) and an assumed density of roughly 1
Mortality:
Up to and including the dose level of 10.0 mL/kg bw no animal died.
In the dose group of 21.5 mL/kg bw 5% (1/20) of the animals was found death two days p.a.. In the dose groups of 31.6 and 46.4 mL/kg bw 88% (14/16) and 100% (20/20) of the animals died within 24 hours or 6 hours p.a. respectively.
Clinical signs:
Especially in the doses 21.5; 31.6 and 46.6 mL/kg quiet, apathetic behavior, closed eyes, diarrhea with soft faeces and decreased sound sensitivity were observed. Also in the 10.0 mL/kg dose group a number of animals showed diarrhea ( occurrence ca. 2 hours after application; this symptom was no longer observed after 1 day after application) . The behavior of the mice in the 1.00 and 4.64 mL/kg was normal active and unremarkable.
Body weight:
No data
Gross pathology:
In the stomach and small intestine of mice who died, dark reddish-brown slimy liquid was determined. The mucous membranes were dark red and translucent, and showed macroscopic signs of irritation, especially the main gastric membranes. Intubation injuries were not found.
Other findings:
- Organ weights: no data
- Histopathology: no data
- Potential target organs: no data
- Other observations: no data

Table 1. Lethality, chronological categorization (summed up, %)

Experimental Period

Dosis (mL/kg)

0 bis

1.00

4.64

10.0

21.5

31.6

46.4

20 min

 

 

 

 

6

 

40 min

 

 

 

 

13

35

60 min

 

 

 

 

19

60

2 h

 

 

 

 

44

70

4 h

 

 

 

 

50

90

6 h

 

 

 

 

 

100

24 h

0

0

0

0

88

100

2 d

 

 

 

5

 

 

3 d

 

 

 

 

 

 

4 d

 

 

 

 

 

 

5 d

 

 

 

 

 

 

6 d

 

 

 

 

 

 

7 d

0

0

0

5

88

100

Number of mice used

10

10

10

20

16

20

Interpretation of results:
GHS criteria not met
Conclusions:
The LD50 at 7 days was 28.7mL/kg. The LD50 for an observation period of 24 hours did practically not differ from the previous value. Recalculated LD50 referring to solid content of 39% (according to producer information) and an assumed density of roughly 1 was calculated to be ca. 11000 mg/kg bw.
Executive summary:

Acute oral toxicity (LD50) of the test item was examined by gavage in male NMRI mice at doses of 1, 4.64, 10, 21.5, 31.6 and 46.4 mL/kg bw. There were no pronounced , specific toxic behavioral symptoms registered at the lower doses of 1 and 4.64 mL/kg bw. At doses ≥10 ml/kg diarrhea was seen. The died mice showed irritation of the mucous membranes of the stomach and small intestine. At the higher doses (21.5, 31.6 and 4.64 mL/kg bw/day), quiet apathetic behaviour, closed eyes and decreased sound activity were also observed.
The LD50
 at 7 days was 28.7 mL/kg. Recalculated LD50 referring to solid content of 39% (according to producer information) and an assumed density of roughly 1 was calculated to be ca. 11000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
11 000 mg/kg bw
Quality of whole database:
Reliable (Klimisch 2)

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
other:
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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: CD/Crl:CD(SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: At dosing: Males: approx. 8 weeks; Females: Approx. 9 weeks
- Weight at study initiation: At dosing: Males: 215 - 237 g; Females: 217 – 239 g
- Fasting period before study: Approx. 16 hours before administration (only tap water was then available ad libitum)
- Housing: During the 14-day observation period the animals were kept in groups of 3 animals in MAKROLON cages ( type III plus) . Granulated textured wood (Granulat A2, J. Brandenburg, 49424 Goldenstedt, Germany)
- 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: July 18, 2012 To: August 6, 2012
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: The shaved intact dorsal skin, between the fore and hind extremities
- % coverage: 5 cm x 6 cm, 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 (Omniplast (P. HARTMANN AG, 89522 Heidenheim, Germany) 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) 4.22 mL/kg bw
- Constant volume or concentration used: yes



Duration of exposure:
24 hours
Doses:
1 dose group (Limit test): 2000 mg/kg bw (dose level refers to the solids ingredients of the test item)
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, 30 and 60 min, as well as 3, 6 and 24 hours after administration. All animals were observed for a period of 14 days. Individual body weights were recorded before administration of the test item and thereafter in weekly intervals up to the end of the study. Changes in weight were calculated and recorded.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs (changes of skin and fur, eyes and mucous membranes, respiratory and the circulatory, autonomic and central nervous system and somatomotor activity as well as behaviour pattern; tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma), body weight, gross pathology
Key result
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:
Under the present test conditions, a single dermal administration of 2000 mg Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1- oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts/kg bw to rats revealed no signs of toxicity.
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:
GHS criteria not met
Conclusions:
According to the EC-Commission directive 67/548/EEC and its subsequent amendments on the approximation of the laws, regulations and administrative provision relating to the classification, packaging and labelling of dangerous substances and the results obtained under the present test conditions,Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1- oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts requires no labeling (as LD50 > 2000 mg/kg).
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, Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1- oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts was examined for acute toxicity after a single dermal application to rats. One dose level of 2000 mg/kg bw was administered on the shaved intact dorsal skin, between the fore and hind extremities of 5 male and 5 female CD/Crl:CD(SD) rats. 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. All animals were observed for a period of 14 days. Under the present test conditions, a single dermal administration of 2000 mg active ingredient/kg bw to rats revealed no signs of toxicity and no deaths. 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:
Reliable (Klimisch 2)

Additional information

Acute oral toxicity

- In a key acute oral toxicity study, the registered substance was examined by gavage in male NMRI mice at doses of 1, 4.64, 10, 21.5, 31.6 and 46.4 mL/kg bw (Hackenberg, 1967). There were no very pronounced, specific toxic behavioral symptoms registered at the lower doses of 1 and 4.64 mL/kg bw. At doses ≥10 mL/kg diarrhea was seen. The died mice showed irritation of the mucous membranes of the stomach and small intestine. At the higher doses (21.5, 31.6 and 4.64 mL/kg bw/day), quiet apathetic behaviour, closed eyes and decreased sound activity were also observed. The LD50 at 7 days was 28.7 mL/kg. Recalculated LD50 referring to solid content of 39% (according to producer information) and an assumed density of roughly 1 was calculated to be ca. 11000 mg act. ingr./kg bw which is above limit dose.

- A supporting oral acute toxicity study in rats with read-across substance ' Butanedioic acid, 2 (or 3) - sulfo-, 4 - [2- [(1-oxo (C12 - C18 (even numbered) and C18unsaturated) alkyl)) amino] ethyl] esters, disodium salts' tested at 2000 mg active ingredient/kg bw did not reveal any signs of toxicity. (Haferkorn, 2013a) No death was recorded within the 14 days observation period. All animals gained the expected weight throughout the whole study period. No pathological changes were observed at necropsy. The LD50value was ranked exceeding 2000 mg/kg bw.

In conclusion there is no hazard for oral acute toxicity with registered substance.

 

Acute dermal toxicity

For the acute dermal toxicity testing, no test data were available for current substance, however read across data were available from 'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts'.

Justification for read across within the N2 (& N3) subgroup of sulfosuccinates is documented in a separate document attached in Section 13. A key dermal toxicity study with that test item containing 41.5% active ingredient was performed at one dose level of 2000 mg active ingredient/kg bw administered on the shaved intact dorsal skin of 5 male and 5 female CD/Crl:CD(SD) rats (Haferkorn, 2013b). The test item was held in contact with the skin under occlusive dressing for 24 hours. All animals were observed for a period of 14 days. The test item revealed no signs of toxicity and no deaths. All animals gained the expected body weight throughout the whole experimental period. No macroscopic findings were observed at necropsy.

In conclusion it can be assumed that there is no hazard for dermal acute toxicity with registered substance.

 

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

- No hazard was identified for acute oral and dermal toxicity based on read-across and registered substances.

- Inhalation toxicity testing was waived based upon the fact that acute inhalation exposure as such is very unlikely.

- Further read-across information in provided in Section 13.

Justification for classification or non-classification

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 acute oral and dermal toxicity.