Reaction mass of sodium hydrogen N,N-dibutyl-10-(sulphonatooxy)octadecanamidate and sodium sulphooctadecanoate

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-05-09 to 2012-06-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

N,N-dibutyloleamide and oleic acid, sulphated, sodium salts

EC Number:

915-926-9

Molecular formula:

C18H34Na2O6S + C26H52NNaO5S

IUPAC Name:

N,N-dibutyloleamide and oleic acid, sulphated, sodium salts

Test material form:

liquid

Details on test material:

The water content of the actual test item was 57.6% (w/w); the water content of the REACH registration substance was analytically determined to be 3.8% (w/w).

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Five male and five female Wistar (RccHan:WIST) strain rats were supplied by Harlan Laboratories UK Ltd., Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non-pregnant. After an acclimatization 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 200 g, 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 wood flakes. 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 (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) 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.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

The calculated volume of test item, as received, was applied as evenly as possible to an area of shorn skin (approximately 10 % of the total body surface area) using a graduated syringe.

Duration of exposure:

24 hours

Doses:

Using available information on the toxicity of the test material, a single group of animals was treated as follows:
Dose Level (mg/kg/bw): 4717* (* - equivalent to 2000 mg active ingredient/kg bodyweight)
Specific Gravity: 1.031
Dose Volume (mL/kg/bw): 4.58
Number of Rats: Male (5) / Female (5)

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: days 0, 7 and 14
- Necropsy of survivors performed: yes

Statistics:

no statistical analysis was performed

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 4 717 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 95 % confidence limits not reported

Mortality:

There were no deaths

Body weight:

other body weight observations

Remarks:

Two males and two females showed bodyweight loss during the first week but expected gain in bodyweight during the second week. The remaining animals 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 to slight oedema, blanching of the skin, light brown discolouration of the epidermis, haemorrhage of dermal capillaries, loss of skin elasticity and flexibility, crust formation, small superficial scattered scabs, hardened light brown coloured scab, scab lifting at edges to reveal dried blood, scab lifting to reveal glossy skin, scab cracking, scab undulating and glossy skin. Adverse reactions prevented accurate evaluation of oedema at the test sites of three males. Adverse reactions prevented accurate evaluation of erythema and/or oedema at the test sites of three females

Any other information on results incl. tables

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.

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
4717*	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

*= Equivalent to 2000 mg active ingredient/kg bodyweight

0= No signs of systemic toxicity

Table 2: 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
4717*	1-0 Male	264	276	304	12	28
	1-1 Male	287	299	334	12	35
	1-2 Male	258	272	297	14	25
	1-3 Male	331	330	352	-1	22
	1-4 Male	385	375	394	-10	19
	2-0 Female	216	217	227	1	10
	2-1 Female	207	202	208	-5	6
	2-2 Female	212	211	216	-1	5
	2-3 Female	200	206	211	6	5
	2-4 Female	202	204	206	2	2

*= Equivalent to 2000 mg active ingredient/kg bodyweight

Table 3: Individual Necropsy Findings

Dose Level mg/kg	Animal Number and Sex	Time of Death	Macroscopic Observations
4717*	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

*= Equivalent to 2000 mg active ingredient/kg bodyweight

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Migrated information: The acute dermal median lethal dose (LD50) of the test item was found to be greater than 4717 mg/kg bodyweight (equivalent to 2000 mg active ingredient/kg bodyweight). 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 4717 mg/kg bodyweight (equivalent to 2000 mg active ingredient/kg bodyweight).

Executive summary:

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” (adopted)

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 4717 mg/kg bodyweight (equivalent to 2000 mg active ingredient/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 to slight oedema, blanching of the skin, light brown discolouration of the epidermis, haemorrhage of dermal capillaries, loss of skin elasticity and flexibility, crust formation, scabbing, scab lifting to reveal dried blood or glossy skin, scab cracking, scab undulating and glossy skin.

Bodyweight: Two males and two females showed bodyweight loss during the first week but expected gain in bodyweight during the second week. The remaining animals 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 4717 mg/kg bodyweight.