N,N-dimethyldecylamine N-oxide

Endpoint summary

Description of key information

Acute oral toxicity: In the key study, performed using C10 AO, the LD50 (rat) was between 300 and 2000 mg AO/kg bw with an LD50 cut-off value of 1000 mg AO/kg bw [Haferkorn J (2012)].

Acute dermal toxicity: One reliable study is available using C10 AO. In this study the reported LD50 (rat) was > 2000 mg AO/kg bw [Haferkorn J (2012)].

Acute inhalation toxicity: No studies are available. The substance is a solid with very low vapour pressure. It is manufactured and supplied as an aqueous solution therefore there is no possibility of exposure to dust. It is used in some spraying applications, but it is expected that inhalation exposure from these applications will be low and hence the dermal route is more relevant.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-11-28 to 2012-02-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

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: ca. 8 weeks
- Weight at study initiation: 160-194 g
- Fasting period before study: yes, ca. 16 hours before start
- Housing: MAKROLON cages (type III plus) with granulated textured wood as bedding.
- Diet (e.g. ad libitum): ad libitum prior to fasting period and in recovery period
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days

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

IN-LIFE DATES: From: 2011-11-28 To: 2011-12-29

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 5.09 mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: based on toxicity information from study sponsor.

Doses:

300 and 2000 mg AO/kg bw

No. of animals per sex per dose:

6 animals at 300 mg AO/kg bw
3 animals at 2000 mg AO/kg bw

Control animals:

no

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 surviving animals were observed for a period of 14 days.
During the follow-up period of two weeks, 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 were observed at least once a day until all symptoms subsided, thereafter each working day. Attention was also paid to possible tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Observations on mortality 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 and at death. Changes in weight were calculated and recorded.

- Necropsy of survivors performed: yes

Statistics:

No applicable

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

300 mg AO/kg bw: No animal died prematurely.
2000 mg AO/kg bw: Two of 3 animals died prematurely within 6 hours after administration.

Clinical signs:

other: Under the present test conditions, a single oral administration of 300 mg N,N-dimethyldecylamine-N-oxide/kg b.w. to female rats revealed slight salivation in 4 of 6 animals. A single oral administration of 2000 mg N,N-dimethyldecylamine-N-oxide/kg b.w. to

Gross pathology:

No pathological changes were observed at necropsy.

Table 1: Summary of results

Symptoms/criteria	300 mg AO/kg bw		2000 mg AO/kg bw
	Females (n = 3) First step	Females (n = 3) Second step	Females (n = 3) First step
Clinical signs: salivation	+ 15’ (3)	+ 5’-15’ (1)	+ 15’ (3)
Mortality: Within 6h Within 24h Within 7d Within 14d
	0	0	2
	0	0	2
	0	0	2
	0	0	2
Mean bodyweight (g) Start After 7d After 14 d
	169.0	171.0	185.0
	195.0 (+15.5)	198.7 (+16.3)	241.0 (+24.1)
	212.7 (+25.9)	220.0 (+28.8)	273.0 (+40.7)
Inhibition of bodyweight gain	none	none	none
Necropsy findings	none	none	none

In brackets:      clinical signs: no of animals affected

                        Bodyweight: bodyweight gain in %, compared to start value

+ = slight

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral (gavage) LD50 is in the range 300 - 2000 mg AO/kg bw.

Executive summary:

Test Guideline

OECD Guideline 423 and EC Method B1 tris (acute toxic class)

Method and material

In a first step 3 female Sprague-Dawley rats were dosed by oral gavage with 300 mg/kg of the test material as supplied (corrected for active content). As no animals died a further three female rats were dosed at the same dose level. No animals died. Three female rats were then dosed by oral gavage at 2000 mg/kg. Animals were observed for 14 days after dosing for mortalities, bodyweight gain and clinical signs of toxicity. All surviving animals under went necropsy at study termination.

Results

300 mg AO/kg bw: No animal died prematurely.

2000 mg AO/kg bw: Two of 3 animals died prematurely within 6 hours after administration.

Salivation was noted in animals in both dose groups after dose administration. There was no effect on bodyweight gain. There were no macroscopic treatment related effects noted at necropsy.

The acute oral (gavage) LD50 is in the range 300 - 2000 mg AO/kg bw.

Conclusion

In accordance with CLP Regulation (EC) No 1272/2008 the substance is classified as Acute category 4 for oral toxicity.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Date started 1986-11-07; Date completed 1986-12-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

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: Interfauna (U.K.) Limited, Wyton, Huntingdon, Cambridgeshire
- Age at study initiation: 5-8 weeks at start of main study
- Weight at study initiation: Males 126-137 g; Females 123-138 g at start of main study
- Fasting period: In the main study, overnight immediately before dosing and for approximately two hours after dosing
- Housing: Solid floor polypropylene cages with sawdust bedding
- Diet: Free access to Rat and Mouse Expanded Diet No. 1, Special Diet Services Limited, Witham, Essex, U.K.
- Water: Free access to mains drinking water
- Acclimation period: Minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 45-65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark controlled by a time switch

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Range-finding study

In order to establish a suitable dose level for the main study a group of two rats (one male and one female) was treated once only. Animals were observed 1 and 4 hours after dosing and then daily for five days.

Main study

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 its fasted bodyweight at the time of dosing. Animals were observed 1 and 4 hours after dosing and subsequently once daily for 14 days. Deaths and evidence of overt toxicity were recorded at each observation.

Doses:

See below

No. of animals per sex per dose:

Range-finding study: 1 male and 1 female

Main study: 5 male and 5 female

Control animals:

no

Details on study design:

The test material was used as supplied. The specific gravity as given by the study sponsor was used to calculate the appropriate dose volumes for the required dose levels. The identity and stability of the test material were not determined.

The animals were selected at random and given a unique number within the study by ear punching and a number written on a cage card.

Range-finding study

Individual bodyweights were recorded on the day of dosing to allow calculation of individual treatment volumes. No necropsies were performed.

Main study

Individual bodyweights were recorded on the day of treatment (day 0) and on days 7 and 14. All animals were subjected to gross necropsy examination for any macroscopic abnormalities. No tissues were retained.

Statistics:

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test material was made. Clinical observations, bodyweights and necropsy records were examined for any adverse but non-lethal effects resulting from treatment.

Preliminary study:

Using the mortality data from the range-finding study (see below), the oral LD50 of the test material was considered to be greater than 2000 mg/kg. This dose level was therefore used for the main study.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: aqueous solution as manufactured and supplied containing 30 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 600 mg/kg bw

Based on:

act. ingr.

Mortality:

No animals died during the range-finding study or the main study (see tables below)

Clinical signs:

other: All treated animlas showed hunched posture and pilo-erection one and four hours after treatment in the main study. All animals recovered and appeared normal on day one and for the rest of the study period. Individual clinical observations are given in Tab

Gross pathology:

Abnormalities noted at necropsy of animals killed at the end of the main study were scattered white raised areas covering approximately 25% of the non-glandular region of the stomach. No other abnormalities were noted. Individual necropsy findings are given in Table 3 (attached).

Other findings:

No data

Mortality data for the range-finding study

Dose Level Mg/kg	Sex	Deaths on Day						Total Deaths
		0	1	2	3	4	5
2000	Male	0	0	0	0	0	0	0/2
	Female	0	0	0	0	0	0

Mortality data for main study

Dose Level m/kg	Sex	Deaths on Day								Total Deaths
		1	2	3	4	5	6	7	8-14
2000	Male	0	0	0	0	0	0	0	0	0/10
	Female	0	0	0	0	0	0	0	0

Conclusions:

The acute oral median lethal dose (LD50) of the test material, a 30% solution of C12-14 AO, to the rat was found to be > 2000 mg/kg bodyweight. This result equates to an LD50 of > 600 mg/kg bodyweight based on active ingredient.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1987-07-16 to 1987-07-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hoechst AG, Kastengrund SPF breeding colony
- Age at study initiation: 6-10 weeks
- Weight at study initiation: 164.0- 173.0 g (males); 162.0-172.0 g (females)
- Fasting period: From about 16 hours before to 3-4 hours after treatment
- Housing: In fully air-conditioned rooms in macrolon cages (type 4) on soft wood granulate in groups of 5 animals
- Diet: Altromin 1324 ad libitum
- Water: Tap water in plastic bottles ad libitum
- Acclimation period: At least seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 50 ± 20
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 hours daily

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The observation period following treatment lasted for 14 days.

Doses:

The animals received the compound as a 20% solution in water, the administration volume being 10 mL/kg body weight

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

The observation period following treatment lasted 14 days. Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During this time the animals were weighed weekly. At the end of the observation period the animals were killed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes.

Statistics:

No data

Preliminary study:

No data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: aqueous solution as manufactured and supplied containing 30 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 600 mg/kg bw

Based on:

act. ingr.

Mortality:

No deaths occurred during the study

Clinical signs:

other: No clinical symptoms were observed after administration of 2000 mg/kg body weight

Gross pathology:

The animals killed at the end of the observation period showed no macroscopically visible changes

Conclusions:

Acute oral toxicity testing of the test item, a 30% aqueous solution of C12-14 AO, in the rat yielded a median lethal dose (LD50) above 2000 mg/kg body weight in both male and female animals. This result equates to an LD50 of >600 mg/kg bodyweight based on active ingredient.

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

The acute oral LD50 in rats was determined using 10 rats (5/sex/group) administered a single oral dose followed by a 14-day observation period.

GLP compliance:

no

Remarks:

Study pre-dates GLP.

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

other: The test material was administered in the form supplied for the highest dose group; all others were diluted with water.

Doses:

15, 5, 0.5, 0.05 and 0.01 g/kg; followed by two further groups dosed at 15.1 and 5.1 g/kg.

No. of animals per sex per dose:

Five rats/sex/group in the definitive test; one rat/sex/group in the pilot study.

Control animals:

no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: aqueous solution as manufactured and supplied containing 6 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 300 mg/kg bw

Based on:

act. ingr.

Conclusions:

The 14-day acute oral toxicity LD50 of the test substance, a 6% aqueous solution of C12-14 AO, in the rat was in excess of 5.0 g/kg bw and would be classed as practically nontoxic according to the toxicity classes of Hodge and Sterner, 1943. There were no deaths or any overt signs of toxicity during the 14-day observation period at this dose. At 15 .1 g/kg bw all ten rats were lethargic within 4 h of treatment. Two/sex died 24 h after treatment and a further 2 male and 1 female rats died 48 h after treatment. The remaining three rats survived the 14 day test period and showed no overt signs of toxicity from Day 3 onwards.

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

10-05-1983 to 24-10-1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

The acute toxicity of the test material over a 14 day period was assessed through administration of a single oral dose to 10 rats (5/sex). The dose was determined by an initial range-finding study.

GLP compliance:

no

Remarks:

Study pre-dates GLP.

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

water

Doses:

25, 200 and 2000 mg/kg bw

No. of animals per sex per dose:

5/sex/group

Control animals:

no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: aqueous solution as manufactured and supplied containing 30 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 600 mg/kg bw

Based on:

act. ingr.

Conclusions:

The 14 day acute oral LD50 of the test substance, a 30% aqeous solution of C12-14 AO, was > 2000 mg/kg bw (equivalent to >600 mg AO/L), the maximum dose tested in this study. There were no deaths and no overt signs of toxicity.

Reference 6

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1977-11-18 - 1977-12-01

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

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

Pre-GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Ind.
- Age at study initiation: Not available
- Weight at study initiation: Male- 211 - 264 gm and Female- 190 - 232 gm
- Fasting period before study: 18-20 hours
- Housing: Individually housed in stainless steel cages with stainless steel grid flooring
- Diet (e.g. ad libitum): Purina Rat Chow manufactured by the Ralston Purina Co.
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: Minimum 4 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.7 - 22.8 °C
- Humidity (%): 46 - 63%
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 12 hours on/off fluorescent lighting


IN-LIFE DATES: From: 1977-11-18 To: 1977-12-01

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Not applicable
- Amount of vehicle (if gavage): Not applicable
- Justification for choice of vehicle: Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable


MAXIMUM DOSE VOLUME APPLIED: 1.4 ml


DOSAGE PREPARATION (if unusual): None


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: None

Doses:

1500, 2100, 3000, 4100, and 5800 mg P7270/kg bw which is equivalent to 420, 588, 840, 1148, and 1624 mg DDAO/kg BW

No. of animals per sex per dose:

5 males and 5 females per dose level

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observation was done at 1/4, 1/2, 1, 2 and 4 hours post-administration and daily thereafter for 14 days. Prefasted body weight was taken and on 14th day.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, mortality and gross pathology

Statistics:

LD50 was calculated using the computer program BLISS 17 (D. J. Finney)

Preliminary study:

Not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

3 800 mg/kg bw

Based on:

test mat.

95% CL:

3 100 - 4 800

Remarks on result:

other: aqueous solution as manufactured and supplied containing 28 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 064 mg/kg bw

Based on:

act. ingr.

95% CL:

868 - 1 344

Mortality:

0/5 males and 0/5 females at 1500 mg/kg
0/5 males and 2/5 females at 2100 mg/kg
1/5 males and 0/5 females at 3000 mg/kg
2/5 males and 4/5 females at 4100 mg/kg
5/5 males and 4/5 females at 5800 mg/kg

Clinical signs:

other: The gross toxic signs observed included decreased motor activity and salivation in all five test groups. Pilo erection was observed in the 4100 and 5800 mg/kg dose groups. Blanching and nasal hemorrhaging were observed in the 2100, 3000, 4100 and 5800 mg/

Gross pathology:

1500 mg/kg (male/female): Not remarkable observation
2100 mg/kg (male): Tan discoloration and pale lungs
(female): Gas and fluid in stomach and intestines
3000 mg/kg (males): Gas and fluid in stomach and intestines
(females): Petechiae on lungs
4100 mg/kg (males): Tan discoloration on lungs, fluid filled stomach and intestines, bright red lungs.
(females): Fluid in stomach, yellow fluid in small intestine, Gas and fluid in stomach and intestine, Fluid filled stomach and intestines and Tan discoloration on lungs.
5800 mg/kg (males): Liver- colored lungs, fluid filled stomach and small intestines, Tan discoloration and petechiae on lungs.
(females): Liver- colored lungs, fluid filled stomach and small intestines, Tan discoloration and petechiae on lungs, gas and fluid in
stomach.

Other findings:

- Organ weights: None
- Histopathology: None
- Potential target organs: None
- Other observations: None

None

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Migrated information

Conclusions:

The LD50 of P7270 is 3800 mg/kg which is equivalent to 1064 mg AO/kg on an active DDAO level. Therefore this test material is classified under OECD GHS Toxicity Category IV.

Executive summary:

The LD50 of dodecyl dimethyl amine oxide is 1064 mg/kg bw based on an active substance level (3800 mg/kg nominal as P7270). Therefore it was concluded that this test substance belongs to OECD GHS Toxicity Category IV.

Reference 7

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

31-01-1978 to 28-02-1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

The acute oral toxicity of the test substance was assessed by single oral doses administered by gavage to female rats. Rats were fasted for 16 h before application of the test substance and for 2 h post appplication. Rats were dosed at 3000 and 5000 mg/kg bw with 10 rats per dose. The rats were observed for 14 days, during which they were weighed weekly. All rats underwent macroscopic assessment .

GLP compliance:

no

Remarks:

Study pre-dates GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: SPF-Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 160 to 178 g (mean weight 170 g).
- Fasting period before study: 16 h
- Housing: In plastic cages with wood shavings.
- Diet: Ad libitum Altomin 1324 (Altomin GMBH).
- Water: Ad libitum

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

No data

Doses:

3000 and 5000 mg/kg body weight

No. of animals per sex per dose:

10 females/dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were weighed weekly.
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs and body weight.

Statistics:

No data

Preliminary study:

Not applicable

Sex:

female

Dose descriptor:

LD50

Effect level:

3 000 - 5 000 mg/kg bw

Based on:

test mat.

Sex:

female

Dose descriptor:

LD50

Effect level:

900 - 1 500 mg/kg bw

Based on:

act. ingr.

Mortality:

At a dose of 5000 mg/kg bw of the test material all 10 animals died within 24 h. At 3000 mg/kg bw of test material one out of 10 rats died.

Clinical signs:

other: Prior to death animals exhibited decreased respiratory rate, narrowed palpebral fissures and were found in a prone position.

Gross pathology:

No macroscopic abnormalities were found in surviving animals. In the animals that died, one was found with the intestinal tract full of the preparation. The other necropsies showed no notable observations.

Other findings:

No data

Results:

Dose (mg/kg)	Concentration (%)	Number of dead animals/ Number of total animals
3000	25	1/10
5000	25	10/10

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral toxicity of the test substance was assessed in female rats from a single oral dose. All rats dosed at 5000 mg/kg bw died within 24 h. One rat exposed to 3000 mg/kg bw test substance died. The acute oral LD50 for female rats was between 3000 and 5000 mg/kg/day bw, equivalent to 900 - 1500 mg AO/kg bw.

Reference 8

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

27-09-1983 to 22-11-1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hoescht AG.
- Age at study initiation:
- Weight at study initiation: Males, mean = 202 g (191 - 219 g ); females, mean = 194 g (185 - 205 g)
- Fasting period before study: 16 h before and 2 h after application.
- Housing: Air-conditioned room in makrolon cages (type 4) on soft wood shavings in groups of 5 animals.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 55 ± 10 %
- Photoperiod (hrs dark / hrs light): 12 h light/12 dark

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25 % w/v

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg

Doses:

1250, 2000, 2500, 3150, 4000 and 5000 mg/kg bw.

No. of animals per sex per dose:

5 females only for groups dosed at 1250, 2000 and 2500 mg/kg.
5/sex/dose for the group dosed at 3150 mg/kg.
5 males only for groups dosed at 4000 and 5000 mg/kg.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were weighed weekly
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs and body weight

Statistics:

The LD50, the 95 % confidence range and the calculation of the probit were determined directly from the death rates (Probit analysis by the method of Lindner and Weber; the confidence limits were calculated according Fieller or Sidak).

Preliminary study:

No data

Sex:

male

Dose descriptor:

LD50

Effect level:

4 120 mg/kg bw

Based on:

test mat.

95% CL:

>= 3 400 - <= 5 010

Sex:

female

Dose descriptor:

LD50

Effect level:

2 820 mg/kg bw

Based on:

test mat.

95% CL:

>= 2 360 - <= 3 540

Sex:

male

Dose descriptor:

LD50

Effect level:

1 236 mg/kg bw

Based on:

act. ingr.

Sex:

female

Dose descriptor:

LD50

Effect level:

846 mg/kg bw

Based on:

act. ingr.

Mortality:

There were deaths in the female rats at 2500 mg/kg (1/5) and at 2500 mg/L (4/5).
Deaths in male rats occurred at 4000 mg/kg (3/5) and 5000 mg/kg (4/5).
Animals died at 1 - 2 days post exposure.

Clinical signs:

other: After 10 minutes after treatment male animals were observed squatting or prone on their stomach or side and a narrowing of the lids. After 30 - 60 miniutes these symptoms were also observed in the females. 30 -60 minutes post application diarrhoea was ob

Gross pathology:

The necropsy of the male animals that died during the study showed the stomach and small intestine filled with yellowish-green liquid and partial whitening of the liver was observed. The necropsy of the dead females showed a reddish liquid-filling the gastrointestinal tract. In two animals the thorax was filled with pink liquid. Some animals showed autolytic changes. Surviving animals were free from visible macroscopic changes.

Other findings:

No data

Clinical signs observed in females:

Time after application	From	0 min	10 min	30 min	1 h	2 h	4 h	1 d	2 d	3 d	4 d
	To	10 min	30 min	60 min	2 h	4 h	6 h				14 d
1250 mg/kg bw: Females
Mortality		0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5
Number of animals with symptoms:
Ruffled fur					5	5	5
No symptoms		5	5	5				5	5	5	5
Necropsy: 5/5 no visible macroscopic abnormalities
2000 mg/kg bw: Females
Mortality		0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5
Number of animals with symptoms:
Squatting				2	2	2	2
Ruffled fur					5	5	5
Subdued						5	5
Diarrhoea					5	5	5
No symptoms		5	5	3				5	5	5	5
Necropsy: 5/5 no visible macroscopic abnormalities
2500 mg/kg bw: Females
Mortality		0/5	0/5	0/5	0/5	0/5	0/5	1/5	1/5	1/5	1/5
Number of animals with symptoms:
Squatting			2	3	5	3	5
Narrow palpebral fissures			3	4	5	5	5
Positioned on their belly or side						2
Belly and flanks drawn in				2	3	5	3
Ruffled fur						5	5
Diarrhoea				2	3	5	5
No symptoms		5						4	4	4	4
Necropsy: 1/5 gastrointestinal tract filled with yellow fluid 1/5 full bulging stomach 4/5 no visible macroscopic abnormalities
3150 mg/kg bw: Females
Mortality		0/5	0/5	0/5	0/5	0/5	1/5	4/5	4/5	4/5	4/5
Number of animals with symptoms:
Squatting				3	3	3
Ruffled fur					5	5
Positioned on their belly or side					2	2	3
Subdued					5	5	4
Decreased respiratory rate						2	4
Diarrhoea			5	5	5	5	5
No symptoms		5						1	1	1	1
Necropsy: 1/5 vessels of the gastrointestinal tract protruding 3/5 gastrointestinal tract filled with reddish fluid 2/5 thorax filled with pink fluid 1/5 gastrointestinal tract filled with yellow fluid 1/5 full bulging stomach 3/5 autolysis (advanced) 1/5 no visible macroscopic abnormalities

Clinical signs observed in males:

Time after application	From	0 min	10 min	30 min	1 h	2 h	4 h	1 d	2 d	3 d	4 d
	To	10 min	30 min	60 min	2 h	4 h	6 h				14 d
3150 mg/kg bw: Males
Mortality		0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5	0/5
Number of animals with symptoms:
Squatting			4	2	2	1	1
Belly and flanks drawn in			5	5	5	5	5
Subdued			5	5	5	5	5
Positioned on their belly or side			1	2	3	3	3
Decreased respiratory rate				1	3	3	3
Ruffled fur				5	5	5	5
High-legged position					2	2	2
Narrow palpebral fissures					4	5	5
Diarrhoea					5	5	5
No symptoms		5						5	5	5	5
Necropsy: 5/5 no visible macroscopic abnormalities
4000 mg/kg bw: Males
Mortality		0/5	0/5	0/5	0/5	0/5	0/5	3/5	3/5	3/5	3/5
Number of animals with symptoms:
Subdued		5	5	5	5	5	5
Narrow palpebral fissures			5	5	3	5	5
Belly and flanks drawn in			5	5	5	3	3
Creeping forward			5	5	5	3	3
Decreased respiration rate				2	2	3	3
Eyelids closed				2	2	2	2
Supine position				1	1
Squatting						2	2
Diarrhoea				3	5	5	5
Strong noisy breathing										2
No symptoms								2	2		2
Necropsy: 2/5 liver partially lightened 1/5 full bulging stomach 3/5 small intestine filled with yellowish-green liquid 3/5 autolysis (early stages) 2/5 no visible macroscopic abnormalities
5000 mg/kg bw: Males
Mortality		0/5	0/5	0/5	0/5	0/5	0/5	2/5	4/5	4/5	4/5
Number of animals with symptoms:
Squatting
Belly and flanks drawn in
Subdued
Supine position
Eyelids closed
Creeping forward
Diarrhoea
Ruffled fur
Noisy breathing
No symptoms
Necropsy: 4/5 liver lightened 4/5 bulging translucent stomach 4/5 small intestine filled with yellowish green liquid 2/5 autolysis (early stages) 1/5 no visible macroscopic abnormalities

Interpretation of results:

Toxicity Category IV

Conclusions:

The acute oral toxicity of the test substance was assessed in male and female rats according to OECD guideline 401 and EU method B.1. The acute oral LD50 of the test substance in males rats was 4120 mg/kg bw, equivalent to 1236 mg AO/kg bw. The acute oral LD50 of the test substance in female rats was 2820 mg/kg, equivalent to 846 mg AO/kg bw..

Reference 9

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

not specified

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

20% dilution

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 per sex/dose

Control animals:

no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 500 mg/kg bw

Based on:

act. ingr.

Mortality:

No mortalities

Clinical signs:

other: None indicated

Interpretation of results:

Toxicity Category IV

Conclusions:

The acute oral median lethal dose (LD50) of the test material to the rat was found to be > 2000 mg/kg bw. This result equates to an LD50 of > 500 mg/kg bw based on active ingredient.

Executive summary:

The acute oral toxicity of C14 AO was assessed in male and female rats in accordance to OECD Guideline No. 401. Five male and five female rats were dosed by oral gavage with 2000 mg/kg bw test material (ca. 25% active). After a 14 day observation period there were no mortalities and no treatment related effects observed on bodyweight or at gross necropsy. The LD50 was > 500 mg AO/kg bw.

Reference 10

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1997-10-29 to 1997-12-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes

Test type:

acute toxic class 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: Harlan Sprague-Dawley, Indianapolis, Indiana
- Age at study initiation: 6 to 10 weeks
- Weight at study initiation: 206 - 260 g
- Fasting period before study: Yes, overnight
- Housing: Individually in stainless steel cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
Temperature, humidity and light controlled room (DHEW publication No. 86-23 (NIH))

IN-LIFE DATES: From: 1997-10-29 To: 1997-11-11

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Dosage volume: 5 g/kg
Metal dosing cannula

Doses:

5 g/kg bw

No. of animals per sex per dose:

5 male/5 female

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Frequently on day of dosing (1, 2.5 and 4 hours) and once daily thereafter
- Necropsy of survivors performed: yes

Statistics:

Calculations of the LC50 and 95% confidence limits: Method of moving averages, Weil CC (1952) table for convenient calculations of median effective dose (LD50 and ED50) and instruction in their use. Biometrics, 8.

Preliminary study:

Not performed

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 1 495 mg/kg bw

Based on:

act. ingr.

Mortality:

No mortalities

Clinical signs:

other: No clinical observations reported

Gross pathology:

No treatment related effects noted at gross necropsy

Interpretation of results:

Toxicity Category IV

Conclusions:

The acute oral administration of the test material resulted in an LD50 >5000 mg/kg bw, equivalent to > 1500 mg AO/kg bw.

Executive summary:

The acute oral toxicity of C14 AO was assessed in male and female rats in accordance to OECD Guideline No. 401. Five male and five female rats were dosed by oral gavage with 5000 mg/kg bw test material (ca. 30% active). After a 14 day observation period there were no mortalities and no treatment related effects observed on bodyweight or at gross necropsy. The LD50 was > 1495 mg AO/kg bw.

Reference 11

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1984-9-13 to 1984-10-18

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: Consumer Product Safety Commission Protocol

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: CD

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River
- Age at study initiation: No data
- Weight at study initiation: 200-300 g
- Fasting period before study: 18 hours
- Housing: raised wire mesh cages
- Diet: Lab Blox ad libitum
- Water: ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%): No data
- Air changes (per hr): Air conditioned quarters
- Photoperiod (hrs dark / hrs light): No data

Route of administration:

other: Rigid stomach tube

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Following administration of each dose level the animals were observed for fourteen days for signs of toxicity.

Doses:

0.4 mL per 100g bodyweight equating to 4.0 g/kg boidyweight
0.5 mL per 100 g bodeweight equating to 5.0 g/kg bodyweight
0.7 mL per 100 g bodyweight equating to 7.0 g/kg bodyweight

No. of animals per sex per dose:

5 males and 5 females per dose level

Control animals:

no

Details on study design:

The animals were examined daily and mortality recorded. An autopsy was performed if autolysis had not occurred.

Statistics:

The method of Litchfield and Wilcoxon was used to calculate the oral LD50 (see Table 1, attached)

Preliminary study:

No data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

5 600 mg/kg bw

Based on:

test mat.

Remarks on result:

other: aqueous solution as manufactured and supplied containing 30 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 680 mg/kg bw

Based on:

act. ingr.

Mortality:

See Table 1 (attached)

Clinical signs:

other: See Table 1 (attached)

Gross pathology:

Autopsy were performed on rats that did not show any signs of autolysis. At autopsy, no macroscopic changes were noted in any of the internal organs.

Other findings:

No data

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The LD50 was calculated to be 5.6 g/kg bodyweight based on test material. This result equates to an LD50 of 1680 mg/kg bodweight based on active ingredient.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

For C10 AO there is one Klimisch score 1 study. This is supported by ten Klimisch score 1 or 2 studies performed on other category members.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 May - 01 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)

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

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: Approximately 8 weeks old
- Weight at study initiation: males 250-262 g; females 208-223 g
- Fasting period before study: 16 h
- Housing: During the 14-day observation period the animals were kept singly in MAKROLON cages (type III plus).
- Diet: Commercial diet, ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany served as food ad libitum. Feeding was discontinued approx. 16 hours before administration
- Water: Drinking water in bottles was offered ad libitum.
- Acclimation period: At least 5 adaptation days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 15
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: From: To: 01 June 2012-26 June 2012

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 5cmx6cm
- % coverage: approximately 10%
- Type of wrap if used: 8 layers of gauze covered with a plastic sheet and secured with adhesive plaster

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not applicable
- Time after start of exposure: not applicable

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5.08 mL/kg bw
- Concentration (if solution): 40.5 % w/w
- Constant volume or concentration used: not applicable
- For solids, paste formed: not applicable

VEHICLE
- Amount(s) applied (volume or weight with unit): Test material supplied as aqueous solution
- Concentration (if solution): 40.5 %w/w
- Lot/batch no. (if required): not applicable
- Purity: not applicable

Duration of exposure:

24 hours

Doses:

2000 mg AO/kg bw

No. of animals per sex per dose:

5 animals/sex/dose

Control animals:

no

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 daily 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, body weight, skin reactions

Statistics:

not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No deaths during the study

Clinical signs:

other: No clinicals signs of toxicity

Gross pathology:

No macroscopic findings were observed at necropsy

Other findings:

skin reactions - no skin reactions were seen in any of the animals

TABLE 1: Summarised results

 

Symptoms/ Criteria	N,N-dimethyldecylamine-N-oxide 2000 mg/kg b.w. (n=5)
	Males	Females
Clinical signs	None	None
Skin reactions	None	None
Mortality Within 6h Within 24 h Within 7d Within 14 d	0 0 0 0	0 0 0 0
Mean body weight (g) Start After 7 days After 14 days	255.4 317.4 (+24.3 %) 370 (+44.9 %)	216.4 235.4 (+8.8 %) 256.0 (+18.3 %)
Inhibition of body weight gain	None	None
Necropsy findings	None	None

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal LD50 is > 2000 mg AO/kg bw

Executive summary:

Test Guideline

OECD Guideline 402 and EC Method B3

Method and material

C10 AO was applied to the shaved backs of CD rats (5 male/5 female) at a dose of 2000 mg AO/kg bw. An occlusive bandage was applied and the rats exposed for a period of 24 hours. After exposure the bandage was removed and the rats observed for 14 days for mortality, clinical signs and skin reactions. At the end of the study all animals were necropsied and examined macroscopically.

Results

There were no deaths and no clinical signs of toxicity. No skin reactions were observed in any of the animals and there were no macroscopic findings at necropsy. Bodyweight gain was normal.

The acute dermal LD50 is > 2000 mg AO/kg bw.

Conclusion

In accordance with CLP Regulation (EC) No 1272/2008 the substance is not classified for acute dernal toxicity.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

21/12/2009 - 19/02/2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

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
- Age at study initiation: males 7 weeks; females 9 weeks
- Weight at study initiation: males 221-235 g; females 206-224 g
- Fasting period before study: 16 hours
- Housing: Housed singly during the 14 day observation period in Makrolon cages
- Diet (e.g. ad libitum): ssniff R/M-H V1534
- Water (e.g. ad libitum): ad libitum
- Acclimation period:at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 55 +/- 15
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: 7/01/2010 - 3/02/2010

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: animals back, 5x6 cm
- % coverage: approx 10 % of body surface
- Type of wrap if used: 8 layers of gauze covered with a plastic sheet and secured with adhesive plaster

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not applicable
- Time after start of exposure: not applicable

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
- Concentration (if solution): not stated
- Constant volume or concentration used: yes

Duration of exposure:

24 hours

Doses:

2000 mg AO/kg bw

No. of animals per sex per dose:

5 animals/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations before and immediately, 5, 15, 30 and 60 min, as well as 3, 6 and 24 hours after administration. During the 14 day observation period observed at least once daily until all symptoms subsided and daily thereafter. Bodyweight recorded before administration and thereafter in weekly intervals up to the end of the study
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Preliminary study:

No preliminary study performed

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No deaths seen during the study

Clinical signs:

other: No clinical signs seen during the study

Gross pathology:

No macroscopic changes were noted at necropsy

Other findings:

- Other observations: Erythema (grade 2-4) was observed at the application site in all 5 males and 5 females immediately after patch removal until test day 6 No oedema was observed in any animals. All reactions cleared by Day 7.

Table 1: Skin reactions observed during the study

Animal No. & sex	Skin reactions on test day
	1	2	3	4	5	6	7
		E/Oe/N	E/Oe/N	E/Oe/N	E/Oe/N	E/Oe/N	E/Oe/N
1 m		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
2 m		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
3 m		2/0/0	0/0/0	3/0/0	4/0/0	4/0/0	0/0/0
4 m		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
5 m		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
6 f		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
7 f		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
8 f		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
9 f		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0
10 f		2/0/0	2/0/0	3/0/0	4/0/0	4/0/0	0/0/0

Animal No. & sex	Skin reactions on test day
	8	9	10	11	12	13	14
		E/Oe/N	E/Oe/N	E/Oe/N	E/Oe/N	E/Oe/N	E/Oe/N
1 m	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
2 m	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
3 m	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
4 m	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
5 m	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
6 f	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
7 f	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
8 f	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
9 f	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0
10 f	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0	0/0/0

m = male         E = erythema   N = necrosis

f = female         Oe = Oedema   0 = no pathological findings

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of C12-18 amine oxide is > 2000 mg AO/kg bw

Executive summary:

The acute dermal toxicity of the substance was investigated in a GLP study performed to OECD 402. The substance was applied as a solution in water to the skin of 5 male and 5 female rats (CD/Crl: CD(SD)) at a dose of 2000 mg AO/kg bw and covered with an occlusive dressing for 24 hours. After completion of the exposure period the dressing was removed. There were no deaths or signs of systemic toxicity during the 14 -day observation period and animals showed the expected bodyweight gains. Erythema (grade 2-4) was observed at the application site in all 5 males and 5 females immediately after patch removal until test day 6. No Oedema was noted at any time period. All reactions cleared after 7 days. Necropsy revealed no macroscopic findings. The LD50 was 2000 mg AO/kg bw.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1978-03-10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Study is comparable to OECD guidelines 402 except that 3 males and 3 female were dosed instead of 5 males and 5 females. Pre-GLP

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

3 males and 3 female were dosed instead of 5 males and 5 females

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

Pre-GLP

Test type:

standard acute method

Limit test:

yes

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kuiper's Rabbit Ranch, Gary, Indiana
- Age at study initiation: Not available
- Weight at study initiation: 2310 - 2810 gm
- Fasting period before study: Not available
- Housing: Individually housed in hanging wire-mesh cages in temperature and humidity controlled quarters
- Diet (e.g. ad libitum): Purina Rabbit Chow, ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): In accordance with the recommendations contained in H.E.W. Publication No. 74-23 (N.I.H.)
- Humidity (%): In accordance with the recommendations contained in H.E.W. Publication No. 74-23 (N.I.H.)
- Air changes (per hr): In accordance with the recommendations contained in H.E.W. Publication No. 74-23 (N.I.H.)
- Photoperiod (hrs dark / hrs light): In accordance with the recommendations contained in H.E.W. Publication No. 74-23 (N.I.H.)


IN-LIFE DATES: Not available

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Back of the animal
- % coverage: 20-30%
- Type of wrap if used: The application site was covered with 8-ply gauze bandaging, rubber dam and several wrappings of 75 mm Elastoplast tape


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Backs were wiped with a wet disposable paper towel
- Time after start of exposure: 24 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2 ml/kg
- Concentration (if solution): Undiluted
- Constant volume or concentration used: yes
- For solids, paste formed: Not applicable


VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable

Duration of exposure:

24 hours

Doses:

2000 mg P7270/kg bw (male/female) which is equivalent to 560 mg active dodecyl dimethyl amine oxide/kg bw

No. of animals per sex per dose:

3 males and 3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations were done at 24 hours and daily thereafter for a total of 14 days. Initial, 7th day and 14th day body weights were taken
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, mortality and gross pathology

Statistics:

None

Preliminary study:

Not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: aqueous solution as manufactured and supplied containing 28 %w/w AO

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 560 mg/kg bw

Based on:

act. ingr.

Mortality:

1/6 rabbits died during the observation period. This death was attributed to pneumonia and was not considered to be compound related.

Clinical signs:

other: Males: 1-14 days: 3/3 animals appeared to be normal Females: 1/3 rabbits showed hypoactivity, decreased limb tone, ataxia, anorexia

Gross pathology:

The rabbit which died during the study period: Stomach, pale green fluid contents; Stomach, white mucoid material adhering to mucosa; Lungs, severe congestion and consolidation; Lungs, white film adhering. The death of the rabbit was attributed to pneumonia.
Rabbits which were sacrificed following 14 days of observation: No gross lesions- 2/3 males and 2/2 females
Kidneys, mottled coloration- 1/3 males

Other findings:

- Organ weights: None
- Histopathology: None
- Potential target organs: None
- Other observations: None

None

Interpretation of results:

study cannot be used for classification

Conclusions:

The LD50 of dodecyl dimethyl amine oxide is >560 mg/kg based (or > 2000 mg/kg bw nominal P7270).

Executive summary:

The LD50 of dodecyl dimethyl amine oxide is > 560 mg AO/kg BW (or >2000 mg/kg bw nominal P7270).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

There is one study (Klimisch score = 1) performed on the substance itself, supported by a study performed using C12-18 AO (Klimisch score =1) and a study performed using C12-14 AO (Klimisch score = 2). The quality of the database is good.

Additional information

Acute oral toxicity:

In the key study [Haferkorn J (2012)] the acute oral toxicity of C10 AO was assessed in Sprague Dawley rats in accordance with OECD TG 423 under GLP, using the commercial product as supplied. Three female rats were dosed by gavage with the equivalent of 300 mg AO/kg bw (approximately 741 mg/kg bw commercial product). There were no deaths in this group during the observation period and a further three females were dosed at the same level. There were no deaths in this group either. Four of the six animals dosed at 300 mg AO/kg bw showed slight salivation up to 15 minutes after dosing. Bodyweight gains were normal and there were no macroscopic findings at necropsy. A further group of three females was dosed with the equivalent of 2000 mg AO/kg bw (approximately 4938 mg/kg bw commercial product). Two of the animals died within six hours of administration. All three animals showed slight salivation up to 15 minutes after dosing. Bodyweight gain in the surviving animal was normal and there were no macroscopic findings at necropsy. The LD50 was between 300 and 2000 mg AO/kg bw. According to the OECD 423 test guideline the LD50 cut-off value was considered to be 1000 mg AO/kg bw.

In addition, supporting studies are available for the following category members:

C14 AO: LD50 (rat) > 1495 mg AO/kg bw (OECD TG 401) [Kukulinski M (1997)]; LD50 (rat) > 500 mg AO/kg bw (OECD TG 401) [Hofmann T (1988)].

C12 -14 AO: LD50 (rat) 1064 mg AO/kg bw (OECD TG 401) [Fulfs JC (1978)]; LD50 (rat) >600 mg AO/kg bw (OECD TG 401) [Jones JR et al (1986)]; LD50 (rat) >600 mg AO/kg bw (OECD TG 401) [Hofmann T (1987)]; LD50 (rat) > 600 mg AO/kg bw (non-guideline study) [Gunn J & Goodwin M (1983)] and LD50 (rat) >300 mg AO/kg bw (non-guideline study) [Gill CRB (1977)].

C14 -16 AO: LD50 (rat) 1680 mg AO/kg bw (Consumer Product Safety Commission protocol) [Bullens P (1984)].

C12-18 AO: LD50 (rat) >900 – 1500 mg AO/kg bw (non-guideline study) [Mayer & Weigand (1978)]; LD50 (rat) 846 – 1236 mg AO/kg bw/day (OECD TG 401) [Rupprich & Weigand (1983)].

Acute dermal toxicity:

In the key study [Haferkorn J (2012)] performed according to OECD TG 402 under GLP, C10 AO was applied as a solution in water to the skin of 5 male and 5 female rats (CD/Crl: CD(SD)) at a dose of 2000 mg AO/kg bw and covered with an occlusive dressing for 24 hours. After completion of the exposure period the dressing was removed. There were no deaths, signs of systemic toxicity or of erythema or oedema during the 14 -day observation period and animals showed the expected bodyweight gains. Necropsy revealed no macroscopic findings. The LD50 was > 2000 mg AO/kg bw.

In a supporting study [Haferkorn J (2010)] performed according to OECD TG 402 under GLP C12-18 AO was applied as a solution in water to the skin of 5 male and 5 female rats (CD/Crl: CD(SD)) at a dose of 2000 mg AO/kg bw and covered with an occlusive dressing for 24 hours. After completion of the exposure period the dressing was removed. There were no deaths or signs of systemic toxicity during the 14 -day observation period and animals showed the expected bodyweight gains. Erythema (grade 2-4) was observed at the application site in all 5 males and 5 females immediately after patch removal until test day 6. No Oedema was observed in any animal at any time period. Erythema was not observed from day 7 onwards (Grade = 0). Necropsy revealed no macroscopic findings. The LD50 was > 2000 mg AO/kg bw.

In a second supporting study, performed according to OECD TG 402 [Dean WP (1978)] C12-14 AO was applied as a solution in water to the skin of 3 male and 3 female rabbits (New Zealand White) at a dose of 2000 mg/kg bw (equivalent to 560 mg AO/kg bw) and covered with an occlusive dressing for 24 hours. One rabbit died during the observation period. This death was attributed to pneumonia and was not considered to be related to treatment. One female rabbit showed hypoactivity, decreased limb tone ataxia and anorexia. Surviving animals showed expected bodyweight gains. Necropsy revealed pale green fluid and white mucoid material adhering to mucosa in the stomach, severe congestion, consolidation and white film adhering in the lungs of the animal that died of pneumonia. In the surviving animals no gross lesions were noted except mottled colouration in the kidneys of one male. The LD50 >2000 mg/kg bw (equivalent to > 560 mg AO/kg bw).

Acute inhalation toxicity:

No data are available for the inhalation route. The substance is a solid with very low vapour pressure. It is manufactured and supplied as an aqueous solution therefore there is no possibility of exposure to dust. It is used in some spraying applications, but it is expected that inhalation exposure from these applications will be low and hence the dermal route is more relevant.

Justification for classification or non-classification

Acute oral toxicity: Classification for pure C10 AO is based on the study performed by Haferkorn J (2012) where the LD50 (rat) was between 300-2000 mg AO/kg bw, with an LD50 cut-off value of 1000 mg AO/kg bw. This result is supported by data available from studies for category members C14 AO, C12 -14 AO, C14 -16 AO and C12 -18 AO. Based on this result the classification of the pure amine oxide is Acute Toxicity 4, with the Hazard Statement H302: Harmful if swallowed.

Acute dermal toxicity: An OECD TG 402 study performed using C10 AO resulted in a LD50 (rat) > 2000 mg AO/kg bw [Haferkorn J (2012)]. This is supported by an OECD TG 402 study performed using category member C12-18 AO which resulted in a LD50 > 2000 mg AO/kg bw [Haferkorn J (2010)] and a limit test performed with C12-14 AO which resulted in a LD50 > 560 mg AO/kg bw [Dean WP (1978)]. On the basis of these three studies, classification for acute dermal toxicity is not required for members of the Amine Oxide category..

Acute inhalation toxicity: No data are available. Testing was not performed as the dermal route was considered to be the most likely route of exposure.