Registration Dossier

Administrative data

Description of key information

Oral LD50 = 2500 mg/kg. Cut-off value based on limited mortality observed in 1/6 animals dosed at 2000 mg/kg. The other 5 animals showed no symptoms of toxicity at all. Dermal LD50 = 5000 mg/kg. Cut-off value based on no mortality or any symptoms of systemic toxicity observed in 10 animals dosed at 2000 mg/kg.
These results confirm expected low acute toxicity based on available information on the separate parts of the salts.

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:
Between 20 September 2011 and 11 October 2011.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Principles of method if other than guideline:
The sequence of dosing may not always follow the Test Guideline as shown in the schematic diagram in attachment 1. It is Company Policy to minimisethe number of animals used on each study in accordance with UK Government Home Office guidelines.
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Female Wistar (RccHan:WIST) strain rats were used.
On receipt the animals were randomly allocated to cages. The animals were nulliparous and non-pregnant. After an acclimatisation period of at least five days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The bodyweights fell within an interval of ±20% of the mean initial bodyweight of the first treated group.

The animals were housed in groups of three in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food was allowed throughout the study.
The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25°C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe.
Doses:
2000 mg/kg
No. of animals per sex per dose:
3 female @ 2000 mg/kg
3 female @ 2000 mg/kg
Control animals:
no
Details on study design:
Procedure
Using available information on the toxicity of the test material, 2000 mg/kg was chosen as the starting dose.

Dose Level Concentration Dose Volume Number of Rats
(mg/kg) (mg/ml) (ml/kg) Female
2000 200 10 3
2000 200 10 3

All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted bodyweight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each group to confirm the survival of the previously dosed animals.
The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for up to fourteen days.
Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment or at death.
At the end of the observation period the surviving animals were killed by cervical dislocation. All animals were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs.
The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Preliminary study:
Not applicable.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 95% confidence limits not reported
Mortality:
One animal was found dead one day after dosing.
Clinical signs:
Signs of systemic toxicity noted in one animal four hours after dosing were pilo-erection, hunched posture and noisy respiration. No signs of systemic toxicity were noted in surviving animals.
Body weight:
The surviving animals showed expected gains in bodyweight over the study period.
Gross pathology:
Abnormalities noted at necropsy of the animal that died during the study were abnormally red lungs, dark liver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed at the end of the study.
Other findings:
None

Evaluation of Data

Data evaluations included the relationship, if any, between the exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made as shown in the schematic diagram in Appendix 1 - (attachment 1).

Table 1              Individual Clinical Observations and Mortality Data

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-2

Female

0

0

0

PHRn

X

 

 

 

 

 

 

 

 

 

 

 

 

 

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0


0=     No signs of systemic toxicity

H =     Hunched posture

P =     Pilo-erection

Rn =   Noisy respiration

X =     Animal dead

Table 2              Individual Bodyweights and Weekly Bodyweight Changes

Dose Level

mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight (g)
at Death

Bodyweight Gain (g) During Week

0

7

14

1

2

2000

1-0 Female

162

178

188

 

16

10

1-1 Female

161

191

198

 

30

7

1-2 Female

165

-

-

147

-

-

2-0 Female

178

190

201

 

12

11

2-1 Female

188

195

216

 

7

21

2-2 Female

164

177

189

 

13

12

Table 3              Individual Necropsy Findings

Dose Level

mg/kg

Animal Number and Sex

Time of Death

Macroscopic Observations

2000

1-0 Female

Killed Day 14

No abnormalities detected

1-1 Female

Killed Day 14

No abnormalities detected

1-2 Female

Found dead Day 1

Lungs: abnormally red

Liver: dark

Kidneys: dark

2-0 Female

Killed Day 14

No abnormalities detected

2-1 Female

Killed Day 14

No abnormalities detected

2-2 Female

Killed Day 14

No abnormalities detected

Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Migrated information: >2000 - 5000 mg/kg
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight, with LD50 cut-off of 2500 mg/kg bodyweight (Globally Harmonised Classification System - Category 5 >2000 - 5000 mg/kg bodyweight).
Executive summary:

Introduction.

The study was performed to assess the acute oral toxicity of the test item following a single oral administration in the Wistar strain rat. The method was designed to be compatible with the following:

OECD Guidelines for the Testing of Chemicals No. 423 “Acute Oral Toxicity – Acute Toxic Class Method” (adopted 17 December 2001) Method B1tris Acute Toxicity (Oral) of Commission Regulation (EC) No. 440/2008

Method.

A group of three fasted females was treated with the test item at a dose level of 2000 mg/kg bodyweight. This was followed by a further group of three fasted females at the same dose level.

The test item was administered orally as asolutioninarachis oil BP. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. 

One animal was found dead one day after dosing.

Clinical Observations. 

Signs of systemic toxicity noted in one animal four hours after dosing were pilo-erection, hunched posture and noisy respiration. No signs of systemic toxicity were noted in surviving animals.

Bodyweight. 

The surviving animals showed expected gains in bodyweight over the study period.

Necropsy. 

Abnormalities noted at necropsy of the animal that died during the study were abnormally red lungs, dark liver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Conclusion. 

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight, with LD50 cut-off of 2500 mg/kg bodyweight (Globally Harmonised Classification System - Category 5 >2000 - 5000 mg/kg bodyweight).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
2 500 mg/kg bw
Quality of whole database:
Adequate GLP study with Klimisch score 1. These results confirm expected low toxicity based on available information on the separate parts of the salt (i.e. ABS and MAPDA).

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:
Between 21 September 2011 and 05 October 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.
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:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Five male and five female Wistar (RccHan:WIST) strain rats were used.
On receipt the animals were randomly allocated to cages. The females were nulliparous and non pregnant. After an acclimatisation period of at least five days the animals were selected at random and given a number unique within the study by indelible ink-marking on the tail and a number written on a cage card. At the start of the study the animals weighed at least 200g, and were eight to twelve weeks of age. The weight variation did not exceed ±20% of the mean weight for each sex.

The animals were housed in suspended solid floor polypropylene cages furnished with woodflakes. The animals were housed individually during the 24 Hour exposure period and in groups of five, by sex, for the remainder of the study. Free access to mains drinking water and food was allowed throughout the study. The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25°C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
On the day before treatment the back and flanks of each animal were clipped free of hair.

Using available information on the toxicity of the test item, a group of five male and five female rats was treated with the test item at a dose level of 2000 mg/kg.
Duration of exposure:
24 hours
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
5 Male @2000 mg/kg
5 Female @2000 mg/kg
Control animals:
not required
Details on study design:
On the day before treatment the back and flanks of each animal were clipped free of hair.
Using available information on the toxicity of the test item, a group of five male and five female rats was treated with the test item at a dose level of 2000 mg/kg.
The appropriate amount of test item was applied as evenly as possible to an area of shorn skin (approximately 10% (50cm2) of the total body surface area) using a graduated syringe. A piece of surgical gauze was placed over the treatment area and semi occluded with a piece of self adhesive bandage. The animals were caged individually for the 24 Hour exposure period. Shortly after dosing the dressings were examined to ensure that they were securely in place.
Due to a calculation error, two animals were given the incorrect dose (0.458g and 0.456g as opposed to the correct dose of 0.456g and 0.424g respectively). Given the discrepancy was negligible and no signs of toxicity were noted in any of the other treated animals, this deviation was considered not to affect the purpose or integrity of the study.
After the 24 Hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item. The animals were returned to group housing for the remainder of the study period.
The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.

After removal of the dressings and subsequently once daily for fourteen days, the test sites were examined for evidence of primary irritation and scored according to the following scale from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.31:

EVALUATION OF SKIN REACTIONS
Erythema and Eschar Formation Value

No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to slight eschar formation (injuries in depth) 4

Oedema Formation

No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4

Any other skin reactions, if present were also recorded.
Individual bodyweights were recorded prior to application of the test item on Day 0 and on Days 7 and 14.

At the end of the study the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.



Statistics:
No statistical analysis was performed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 95% confidence limits not reported.
Mortality:
There were no deaths.

Clinical signs:
There were no signs of systemic toxicity.
Body weight:
Two females showed bodyweight loss during the first week with no gain in bodyweight or bodyweight loss the second week. Two females showed bodyweight loss during the first week with expected gains in bodyweight during the second week and the remaining female showed expected gains in bodyweight during the first week but bodyweight loss during the second week. All males showed expected gains in bodyweight over the study period.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
Dermal Reactions

Signs of dermal irritation noted were very slight to well defined erythema, very slight oedema, blanching of the skin, loss of skin elasticity, haemorrhage of dermal capillaries, light brown discolouration of the epidermis, glossy skin, small superficial scattered scabs, hardened light brown coloured scab, scab lifting at edges to reveal dried blood, scab lifting to reveal glossy skin, scab undulating and scab cracking. Adverse reactions prevented accurate evaluation of erythema and oedema at the test sites of all animals.

Evaluation of Data

Data evaluations included the relationship, if any, between the exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.

Using the mortality data obtained, an estimate of the acute dermal median lethal dose (LD50) of the test item was made.

The results were also interpreted according to EU labelling regulations Commission Directive 2001/59/EC and Regulation (EC) No 1272/2008 for the classification, packaging and labelling of dangerous substances.

Table 1              Individual Clinical Observations and Mortality Data

Dose Level

mg/kg

Animal Number and Sex

Effects Noted After Initiation of Exposure (Hours)

Effects Noted After Initiation of Exposure (Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-2

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-3

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-4

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-4

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0


0= No signs of systemic toxicity

Table 4              Individual Bodyweights and Weekly Bodyweight Changes

Dose Level mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Change (g) During Week

0

7

14

1

2

2000

1-0 Male

266

276

313

10

37

1-1 Male

299

308

340

9

32

1-2 Male

256

259

282

3

23

1-3 Male

277

284

306

7

22

1-4 Male

258

268

291

10

23

2-0 Female

231

224

224

-7

0

2-1 Female

225

210

215

-15

5

2-2 Female

228

225

230

-3

5

2-3 Female

212

215

207

3

-8

2-4 Female

224

220

216

-4

-4

Table 5              Individual Necropsy Findings

Dose Level

mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

1-0

Male

Killed Day 14

No abnormalities detected

1-1

Male

Killed Day 14

No abnormalities detected

1-2

Male

Killed Day 14

No abnormalities detected

1-3

Male

Killed Day 14

No abnormalities detected

1-4

Male

Killed Day 14

No abnormalities detected

2-0

Female

Killed Day 14

No abnormalities detected

2-1

Female

Killed Day 14

No abnormalities detected

2-2

Female

Killed Day 14

No abnormalities detected

2-3

Female

Killed Day 14

No abnormalities detected

2-4

Female

Killed Day 14

No abnormalities detected

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.
The test item did not meet the criteria for classification according to EU labelling regulations Commission Directive 2001/59/EC or Regulation (EC) No 1272/2008.
Executive summary:

Introduction. 

The study was performed to assess the acute dermal toxicity of the test item in the Wistar strain rat. The method was designed to be compatible with the following:

OECD Guidelines for the Testing of Chemicals No. 402 “Acute Dermal Toxicity” (adopted24 February 1987)

Method B3 Acute Toxicity (Dermal) of Commission Regulation (EC) No. 440/2008

Method. 

A group of ten animals (five males and five females) was given a single, 24 hour, semi‑occluded dermal application of the undiluted test item to intact skin at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. 

There were no deaths.

Clinical Observations. 

There were no signs of systemic toxicity.

Dermal Irritation. 

Signs of dermal irritation noted were very slight to well‑defined erythema, very slight oedema, scabbing, blanching of the skin, loss of skin elasticity, haemorrhage of dermal capillaries, light brown discolouration of the epidermis, glossy skin, scab lifting to reveal dried blood or glossy skin, scab undulating and scab cracking. 

Bodyweight. 

All females showed no gain in bodyweight and/or bodyweight loss during the study. All males showed expected gains in bodyweight over the study period.

Necropsy. 

No abnormalities were noted at necropsy.

Conclusion. 

The acute dermal median lethal dose (LD50) of the test item in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.

The test item did not meet the criteria for classification according to EU labelling regulations Commission Directive 2001/59/EC or Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
5 000 mg/kg bw
Quality of whole database:
Adequate GLP study with Klimisch score 1. These results confirm expected low toxicity based on available information on the separate parts of the salt (i.e. ABS and MAPDA).

Additional information

The obtained results confirm expected low toxicity based on available information on the sepearte parts of the salts (i.e. sec-alkylbenzene sulfonate (ABS) and 3-(methacrylamidopropyl) dimethylammonium (MAPDA)):

Both ABS and MAPDA are of low acute toxicity with oral LD50 values in 300 to 2000 mg/kg range for ABS and even higher for the methacrylamide part. By dermal route toxicity is possibly even lower. The substance MAPDA-ABS salt, as well as both ABS and MAPDA have (very) low vapour pressures. Due to the low vp (< 0.00000084 Pa), exposure is only possible as aerosols, and in view of general low toxicity, inhalation hazards are also likely limited.

MAPDA-ABS salt contains about two-third of ABS which is considered to be driving the toxicity of the products, as toxicty of MAPDA lower. The results obtained from testing for acute oral and dermal toxicity on MAPDA-ABS salt are in line with this.

Acute toxicity oral:

LD50 value is cut-off value based on limited mortality observed in 1/6 animals dosed at 2000 mg/kg. The other 5 animals showed no symptoms of toxicity at all. These results confirm expected low toxicity based on available information on the separate parts of the salts.

Acute toxicity dermal:

Limit test. No mortality or any symptoms of systemic toxicity was observed in 10 animals dosed 2000 mg/kg. LD50 cut-off is set at 5000 mg/kg.

Acute toxicity inhalation:

The likelihood for exposure via inhalation and thus absorbing enough to cause systemic toxicity, is very low, based on the high boiling point (> 300 °C) and very low vapour pressure (< 0.00000084 Pa at 20°C).

The substance is a salt, and thus does not have specific aliphatic, alicyclic and aromatic hydrocarbon properties. No classification for aspiration is therefore considered.


Justification for selection of acute toxicity – oral endpoint
Only study available.

Justification for selection of acute toxicity – dermal endpoint
Only study available.

Justification for classification or non-classification

Available information indicates low acute toxicity via oral and dermal route with LD50 > 2000 mg/kg bw. For dermal route no classification is needed for GHS. For oral route the cut-off LD50 of 2500 mg/kg leads to GHS Cat. 5 classification, but in EU CLP this does not lead to a classification.

Therefore no classification is needed.