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Description of key information

LD50  (rat, oral) = 3020 (females) or 2950 (genders combined) mg/kg bw
LD50  (rat, dermal) > 2000 mg/kg(expressed as mg active ingredient)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
25 May 1995 - 31 July 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
yes
Remarks:
2000 mg/kg bw of test solution administered, equivalent to 830 mg active ingredient/kg bw (41.5%)
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, supplied by Criffa, S.A., Barcelona, Spain
- Age at study initiation: 4 weeks upon arrival
- Weight at study initiation: 107-122 g upon administration
- Fasting period before study: Food removed approx. 18 hours before administration, until approx. 3 hours after
- Housing: in makrolon cages (55 x 32.7 x 19 cm) with sawdust bedding (replaced by wire floor during fasting period), 5 rats of the same sex/cage
- Diet: Standard rat diet UAR A04C ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 (occasionally reaching 26°C)
- Humidity (%): 40-70 (occasionally reaching 32-39% or 71-86%)
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): 12 / 12 (07.00 am-07.00 pm)

IN-LIFE DATES: From: june 21st, 1995 To: July 14th, 1995
Route of administration:
oral: gavage
Vehicle:
other: bidistilled water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg Betadet SHR/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: Not provided

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw
Doses:
2000 mg test solution/kg bw
No. of animals per sex per dose:
1 female (preliminary assay)
5 males + 5 females (main assay)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: frequently during the day of administration, at least twice a day for the remainder of the observation period
- Frequency of weighing: on days 0 (administration), 1, 2, 3, 7, and 14 (at necropsy)
- Necropsy of survivors performed: yes (main study)
- Other examinations performed: clinical signs, body weight
Statistics:
Not included
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks:
Betadet SHR
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 830 mg/kg bw
Based on:
act. ingr.
Remarks:
based on 41.5% purity
Mortality:
No mortality observed.
Clinical signs:
other: Slightly soft feces noted among treated animals on the day of dosing. No other clinical signs observed for the remainder of the observation period.
Gross pathology:
No macroscopic abnormalities observed.

Dose level (mg/kg)

Animals

Observation day

(Day 0 = day of administration)

Bodyweight at necropsy

No.

Gender

0

1

2

3

7

14

2000

2

M

122

145

160

169

212

282

282

3

M

116

140

154

162

203

270

270

4

M

116

138

154

159

202

260

260

5

M

107

124

141

147

190

256

256

6

M

125

149

164

173

216

284

284

7

F

117

137

148

154

175

211

211

8

F

120

140

149

156

172

200

200

9

F

113

133

142

150

167

193

193

10

F

109

128

142

150

166

194

194

11

F

122

148

158

164

179

207

207

 

Bodyweight measurements (in grams, main study)

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The oral LD50 in rats was higher than 2000 mg Betadet SHR/kg bw, equivalent to approximately 830 mg active ingredient/kg bw.
Executive summary:

Cocamidopropyl hydroxysultaine, as a 41.5% solution, has been tested for acute oral toxicity in Wistar rats. The test article was administered as a single oral dose by gavage. One group of 5 rats per sex received a dose of 2000 mg test solution/kg bw. Examinations for mortality and clinical signs were performed daily during the 14 -day study period. Animals were necropsied at the end of the observation period.

 

No mortality was observed. Slightly soft feces were noted among treated animals on the day of dosing, but no other clinical signs were observed over the rest of the observation period. Bodyweight was not affected by the administration. No relevant changes were seen at necropsy.

 

The oral LD50 in rats was therefore higher than 2000 mg test solution/kg bw, equivalent to approximately 830 mg active ingredient/kg bw.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
other: OECD test guideline 401 ‘Acute Oral Toxicity’ deleted on 17th December 2002
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany
- Age at study initiation: not provided
- Weight at study initiation: 171-276 g (males), 164-216 g (females)
- Fasting period before study: from 16 hours before until 3-4 hours after administration
- Housing: up to a maximum of 5 rats per cage (Macrolon type III cage)
- Diet (e.g. ad libitum): not detailed
- Water (e.g. ad libitum): not detailed
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 2
- Humidity (%): 50-85
- Air changes (per hr): not detailed
- Photoperiod (hrs dark / hrs light): 12 / 12 (7.00 am-7.00 pm)

IN-LIFE DATES: Not provided
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
No vehicle used (solution administered as such with different dosing volumes)

MAXIMUM DOSE VOLUME APPLIED:
1000 mg/kg: 2.33 mL/kg bw
2000 mg/kg: 4.65 mL/kg bw
3000 mg/kg: 6.98 mL/kg bw
Doses:
1000, 2000 and 3000 mg active component/ kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Examinations performed: mortality (over the 24-hour post-dosing period, and then daily), clinical signs (daily), body weights (just before dosing, on days 7 and 14)
- Necropsy of survivors performed: yes
Statistics:
The method of Finney D.Y., Probit Analysis (3rd ed., Cambridge, 1971) was used for calculating the oral LD50.
Preliminary study:
One animal died within 24 hours of dosing at 2000 mg/kg
Sex:
male
Dose descriptor:
LD50
Remarks on result:
other: Could not be calculated because only at the high dose pre-terminal deaths were lower than 100% and higher than 0%
Sex:
female
Dose descriptor:
LD50
Effect level:
3 020 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: at 24 h and 14 days after dosing
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 950 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: at 24 h and 14 days after dosing
Mortality:
See table below
Clinical signs:
other: Up to 3 days post-dosing, reduced general activity was observed at 3000 mg/kg, together with squatting position, reduced skin turgor, cyanosis, diarrhea and piloerection on some occasions.
Gross pathology:
- 2000 and 3000 mg/kg: animals killed in extremis within 24 hours post-dosing showed hemorrhagic and lytic alterations in the gatro-intestinal tract and/or yellow-orange discoloration of lungs and/or reddish pelvis at macroscopic examination.
- At terminal sacrifice (14 days post-dosing): no test-article abnormalities noted at necropsy.

Dose (mg/kg)

Post-treatment time

Males

Females

24 hours

7 days

14 days

24 hours

7 days

14 days

1000

0/5

0/5

0/5

0/5

0/5

0/5

2000

0/5

0/5

0/5

2/5

2/5

2/5

3000

3/5

3/5

3/5

2/5

2/5

2/5

Cumulative Mortality over the 14-day observation period

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The oral LD50 in rats was calculated to be 3020 or 2950 mg active ingredient/kg bw, for females or both genders, respectively, at 24 hours and 14 days after administration. The oral LD50 in male rats could not be calculated because deaths occurred only in the high-dose group.
Executive summary:

Cocamidopropyl hydroxysultaine, as a 42% aqueous solution, has been tested for acute oral toxicity in Wistar rats. The test article was administered as such using different dosing volumes to reach the desired dose levels. Three groups of 5 rats per gender received a dose volume of 2.33, 4.65 and 6.98 mL/kg, equivalent to 1000, 2000 and 3000 mg active ingredient/kg, respectively. Examinations for mortality and clinical signs were performed daily during the 14-day study period. Body weights were measured just before dosing, and 7 and 14 after dosing. A macroscopic examination was performed at the necropsy of survivors on day 14.

 

A high incidence of pre-terminal deaths occurred at 2000 and 3000 mg/kg, whereas no death occurred at 1000 mg/kg: 3/5 male rats were found dead or sacrificed in extremis within 24 hours of dosing in the 3000 mg/kg dose group, and 2/5 female rats were found dead or sacrificed in extremis within 24 hours of dosing in each of the 2000 mg/kg and 3000 mg/kg dose groups. Marked clinical signs, such as general reduced activity together with diarrhea, squatting position, piloerection and/or reduced skin turgor were observed at 3000 mg/kg within 3 days post-dosing. Body weight gain was normal in surviving animals over the observation period. Hemorrhagic and lytic mucous membrane alterations in the gastro-intestinal tract, considered test-article related, were observed at necropsy in animals found dead or sacrificed in extremis within 24 hours of dosing in the 2000 mg/kg and 3000 mg/kg dose groups. At terminal sacrifice, no test-related macroscopic findings were observed in the other animals.

The oral LD50 in rats was calculated to be 3020 or 2950 mg active ingredient/kg bw, for females or both genders, respectively, at 24 hours and 14 days after administration. The oral LD50 in male rats could not be calculated because deaths occurred only in the high-dose group.

Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
C8-C18 AAPHS and C12 AAPHS have the same functional groups, and general composition. The main variable resides in the alkyl chain distribution.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source chemical = C8-18 cocamidopropyl hydroxysultaine (EC 939-455-3)
Target chemical = C12 cocamidopropyl hydroxysultaine (EC 242-893-1)

3. ANALOGUE APPROACH JUSTIFICATION
The alkyl C-chain distribution of the source chemical significantly overlaps with the one of the target chemical, the C12-alkyl derivative being the major constituent in both chemicals. The structural differences in side C-chains is not expected to lead to significant differences on phys-chem properties.

4. DATA MATRIX: see "Documentation and scientific justification of the read-across approach" in section 13.2.
Reason / purpose for cross-reference:
read-across source
Sex:
male
Dose descriptor:
LD50
Remarks on result:
other: Could not be calculated because only at the high dose pre-terminal deaths were lower than 100% and higher than 0%
Sex:
female
Dose descriptor:
LD50
Effect level:
3 020 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: at 24 h and 14 days after dosing
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 950 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: at 24 h and 14 days after dosing
Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The oral LD50 in rats was calculated to be 3020 or 2950 mg active ingredient/kg bw, for females or both genders, respectively, at 24 hours and 14 days after administration. The oral LD50 in male rats could not be calculated because deaths occurred only in the high-dose group.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
2 950 mg/kg bw
Quality of whole database:
Two Klimisch score 2 studies in rat are available, with consistent study results

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
February - August 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:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approx. 8 weeks old
- Weight at study initiation: 320-348 g (males) - 214-248 g (females)
- Fasting period before study: No
- Housing: By 5 from the same sex and group in polycarbonate cages with stainless steel lids containing autoclaved sawdust, with nylabone used as enrichment
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 8 days (males) - 5 days (females) before application

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 50 +/- 20
- Air changes (per hr): 12 cycles
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 05 March 2012 To: 23 March 2012
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: approx. 7 x 5 cm for males, 6 x 5 cm for females
- % coverage: approx. 10% of total body surface
- Type of wrap if used: Application site covered with aerated hypoallergic dressing

REMOVAL OF TEST SUBSTANCE
No washing

TEST MATERIAL
- Solution applied as is.
- Quantity of test item applied adjusted based on the body weight recorded on the day of application.
- Correction factor of 2.76 used to calculate the dosage volume to be applied taking account of the solution purity.
Duration of exposure:
24 hours
Doses:
2000 mg active ingredient/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
. Morbidity / mortality: frequently during hours following application, at least once daily for the remainder of the observation period
. Clinical signs: at least once during first 30 minutes, periodically during first 4 hours, once daily for the remainder of the observation period
. Bodyweight: recorded on the day of group allocation, then on the day of application (day 1) and on days 8 and 15
- Necropsy of survivors performed: yes (spleen was preserved in 10% buffered formalin, stored, and destroyed at finalization of study report)
Statistics:
No statistical analyses included
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
act. ingr.
Mortality:
No unscheduled deaths observed
Clinical signs:
other: No clinical signs indicative of systemic toxicity observed. Very slight or well defined erythema noted at application site for 2 females on day 2.
Gross pathology:
Enlargement of the spleen was found in all treated animals but was considered incidental as it is part of the normal background in untreated rats of these strain and age.
Other findings:
Due to enlargement seen at necropsy, spleens were preserved in 10% buffered formalin and stored. In the absence of toxicological relevance, no histopathology was conducted and these organs were therefore destroyed upon finalization of the study report.

 

Females

Males

Historical controls

Treated

Historical controls

Treated

Dose level (mg/kg)

0

2000

0

2000

Days 1-8

+36

+21

+45

+41

Days 8-15

+18

+16

+45

+44

Days 1-15

+55

+37

+90

+84

 

Mean bodyweight gains (grams) during the observation period

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 in rats was higher than 2000 mg active ingredient/kg bw.
Executive summary:

Cocamidopropyl hydroxysultaine, as a 36.2% solution (Mackam CBS-50GE), has been tested for acute dermal toxicity in Sprague-Dawley rats. The test article was applied as a single dose under a semi-occlusive dressing for 24 hours. One group of 5 rats per sex received a dose of 2000 mg active ingredient/kg bw. Examinations for mortality and clinical signs were performed daily during the 14 -day study period. Animals were necropsied at the end of the observation period.

 

No mortality was observed. No clinical signs indicative of systemic toxicity were observed. Very slight or well defined erythema was noted at the application site for 2 females on day 2. Mean bodyweight gain was slightly lower than historical control data for females over the observation period, notably during the first week following application. However, no such changes were observed for males. Enlargement of the spleen was found in all treated animals but was considered incidental as it is part of the normal background in untreated rats of these strain and age.

The dermal LD50 in rats was therefore higher than 2000 mg active ingredient/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
C8-C18 AAPHS and C12 AAPHS have the same functional groups, and general composition. The main variable resides in the alkyl chain distribution.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source chemical = C8-18 cocamidopropyl hydroxysultaine (EC 939-455-3)
Target chemical = C12 cocamidopropyl hydroxysultaine (EC 242-893-1)

3. ANALOGUE APPROACH JUSTIFICATION
The alkyl C-chain distribution of the source chemical significantly overlaps with the one of the target chemical, the C12-alkyl derivative being the major constituent in both chemicals. The structural differences in side C-chains is not expected to lead to significant differences on phys-chem properties.

4. DATA MATRIX: see "Documentation and scientific justification of the read-across approach" in section 13.2
Reason / purpose for cross-reference:
read-across source
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
act. ingr.
Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 in rats exposed to C8-C18 Cocamidopropyl dimethyl hydroxysultaine was higher than 2000 mg active ingredient/kg bw. By analogy, the same conclusion applies to C12-C18 Cocamidopropyl dimethyl hydroxysultaine.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
Recent GLP and OECD test guideline-compliant study (Klimisch score 1)

Additional information

In the absence of studies conducted according to OECD guidelines, a read-across was therefore performed with the cocamidopropyl hydroxysultaine which contains alkylamidopropylhydroxysultaine with carbon chain from C8 to C18. The main component of Cocamidopropylhydroxysultaine C8-18 is lauramidopropylhydroxysultaine. Justification for the read-across is documented in a separate document attached in Iuclid Section 13.

Based on the nature of the substance and its likely routes of exposure, the acute toxicity of Cocamidopropyl hydroxysultaine has been tested by oral and dermal routes.

Two Klimisch score 2 studies in rat were available for oral route. One study was used as a key study and the other two as supporting:

 

  • In the key study (1990), Cocamidopropyl hydroxysultaine was administered by gavage at 1000, 2000 or 3000 mg active ingredient/kg as a 42% active ingredient aqueous solution to 5 rats per gender, kept for a 14-day observation period and then necropsied. A high incidence of pre-terminal deaths occurred at 2000 and 3000 mg/kg, whereas no death occurred at 1000 mg/kg. Marked clinical signs were observed at 3000 mg/kg within 3 days post-dosing. Hemorrhagic and lytic mucous membrane alterations in the gastro-intestinal tract, considered test-article related, were observed at necropsy in animals found dead or sacrificed in extremis within 24 hours of dosing in the 2000 mg/kg and 3000 mg/kg dose groups. The LD50 was calculated to be 3020 or 2950 mg active ingredient/kg bw, for females or both genders, respectively. 
  • In the supporting rat study (1995), Cocamidopropyl hydroxysultaine was administered by gavage at the dose of 2000 mg test solution/kg as a 41.5% active ingredient aqueous solution to 5 animals per gender, kept for a 14-day observation period and then necropsied. No mortality, overt sign of toxicity, or relevant bodyweight changes were observed. Therefore, the LD50 was higher than 2000 mg test solution/kg, equivalent to 830 mg active ingredient/kg.

One Klimisch score 1 study was available for dermal route and was used as a key study. In this study (2012), Cocamidopropyl hydroxysultaine was applied for 24 hours under semi-occlusive coverage at the dose of 2000 mg active ingredient/kg as a 36.2% active ingredient aqueous solution to 5 rats per gender, kept for a 14-day observation period and then necropsied. No mortality, overt sign of toxicity, or relevant bodyweight changes were observed. Therefore, the LD50 was higher than 2000 mg active ingredient/kg. Based on the absence of mortality or severe clinical signs in rats up to 2000 mg active ingredient/kg, no classification for dermal acute toxicity is warranted. No target organ was identified following a single dermal application in rats.


Justification for selection of acute toxicity – oral endpoint
Study with the highest degree of compliance with presently recommended test standards.

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

Justification for classification or non-classification

Based on the LD50 values obtained in the key study performed on the analogue substance Cocamidopropyl hydroxysultaine, no classification for oral acute toxicity is warranted according to Reg. (EC) No 1272/2008 (CLP) criteria. This is supported by the absence of mortality or severe clinical signs in rats up to 830 mg active ingredient/kg seen in the other study under consideration.

In accordance with CLP Regulation (EC) No 1272/2008 criteria, as the dermal LD50 in rats is higher than 2000 mg active ingredient/kg, no classification for dermal acute toxicity is warranted.

No target organ was identified following a single oral or dermal administration in rats.