Fatty acids, C18-unsatd., reaction products with acrylic acid and polyethylenepolyamines

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study was conducted in accordance with international guidelines and in accordance with GLP. All relevant validity criteria were fulfilled.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Monitoring authority

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C18-unsatd., reaction products with acrylic acid and polyethylenepolyamines

EC Number:

287-820-4

EC Name:

Fatty acids, C18-unsatd., reaction products with acrylic acid and polyethylenepolyamines

Cas Number:

85586-18-1

Molecular formula:

Not applicable (a generic molecular formula cannot be provided for this specific UVCB substance)

IUPAC Name:

(Z,9Z)-N-(2-{2-[(8Z)-heptadec-8-en-1-yl]-4,5-dihydro-1H-imidazol-1-yl}ethyl)octadec-9-enimidic acid; 3-[(2-carboxyethyl)({2-[3-(2-carboxyethyl)-2-[(8Z)-heptadec-8-en-1-yl]imidazolidin-1-yl]ethyl})amino]propanoic acid; 3-{2-[(8Z)-heptadec-8-en-1-yl]-3-{2-[(E)-[(9Z)-1-hydroxyoctadec-9-en-1-ylidene]amino]ethyl}imidazolidin-1-yl}propanoic acid

Test material form:

other: liquid

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 (HSdlHAN: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 acclimatisation period of at least five days, the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on the cage card. At the start of the study the animals weighed at least 200 g, and were eight to twelve weeks of age.

The animals were housed in suspended solid floor polypropylene cages furnished with woodflakes. The animals were housed individually during the 24 hour exposure period and gang housed up to four per sex for the remainder of the study. Free access to mains water and food was allowed thoughout the study. Food, water and bedding are routinely anlaysed by the laboratory 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 humidity were set to within limits of 19 to 25 degrees centigrade and 30 to 70 percent relative humidity respectively. The rate of air exchange was at least fifteeen changes per hour. Lighting was controlled to a twelve hour light (06:00 am to 18:00 pm) dark cycle.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

On the day before treatment the back and flanks of each animal were clipped free of hair. Initially one male and one female were treated at a dose level of 2,000 mg/kg in the absence of data suggesting any toxicity.

The calculated volume of test material as received was applied evenly to an area of shorn skin (approximately 10% of the total body surface area using a graduated syringe. A piece of surgical gauze was placed over the treatment area and semi occluded with a piece of self-adhesive tape. The animals were caged individually for the 24 hour exposure period . Shortly after dosing the dressings were examined to ensure that they were securely in place.

After the 24 hour contact period, the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with distilled water to remove ant residual test material.

As no mortalities were noted a further group of animals (four male and four females) was similiarly treated with the test material at a dose of 2,000 mg/kg body weight to give a total of five male and five females. After the 24-hour contact period the bandages were carefully removed, the skin and surrounding hair cleaned with cotton wool moistened with distilled water. These animals were gang housed for the remainder of the study.

Duration of exposure:

24 hour exposure period with fourteen day post exposure period.

Doses:

2,000 mg/kg

No. of animals per sex per dose:

Five

Control animals:

not required

Details on study design:

The animals were observed for death or overt signs of toxicity 30 minutes, one hour, two hours and four hours after dosing and subsequently once daily for fourteen days.

After removal of the dressings and subsequently once daily for fourteen days, the test sites were examined for evidence of primary irritation and scored according to the Draize scale (Draize JH (1977) Dermal and eye toxicity tests). Any other skin reactions if present were also recorded.

Individual body weights were recorded prior to the application of the test material on Day 0, Day 7 and 14.

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

Statistics:

None

Results and discussion

Preliminary study:

LD50 > 2,000 mgKg

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths in either sex

Clinical signs:

other: There were no signs of systemic toxicity in either sex

Gross pathology:

Individual examinations were made
No abnormalities were noted

Other findings:

Individual dermal reactions were measured.
Very slight to well defined erythema was noted at the test sites of four males and four females. Other dermal reactions noted at the test site of one female was haemorrhage of dermal capillaries, small superfical scattered scabs and crust formation. Treated skin sites of one male and one female appeared normal throughout the study. The remaining treated skin sites of all other animals appeared normal 3 to 9 days after dosing.

Any other information on results incl. tables

Individual Clinical observations and mortality data

Dose mg/kg	Animal & Sex	1/2hr	1hr	2hr	4hr	1d	2d	3d	4d	5d	6d	7d	8d	9d	10d	11d	12d	13d	14d
2000	1-0 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	3-0 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	3-1 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	3-2 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	3-3 Male	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	2-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	4-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	4-1 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	4-2 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2000	4-3 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Individual Bodyweights and weekly bodyweight changes

Dose mg/kg	Animal & Sex	BW day 0 gm	BW day 7 gm	BW day 14 gm	BW Change gm during week 1	BW Change gm during week 2
2000	1-0 Male	245	263	280	18	17
2000	3-0 Male	237	254	302	17	48
2000	3-1 Male	238	269	298	31	29
2000	3-2 Male	231	252	281	21	29
2000	3 -3 Male	231	252	281	21	29
2000	2-0 Female	211	212	220	1	8
2000	4-0 Female	216	218	222	2	4
2000	4-1 Female	216	215	218	-1	3
2000	4-2 Female	219	220	221	1	1
2000	4-3 Female	203	203	206	0	3

Where BW = Bodyweight

Individual necropsy findings

Dose Mg/kg	Animal & Sex	Time of death	Macroscopic observations
2000	1-0 Male	Killed day 14	No abnormalities detected
2000	3-0 Male	Killed day 14	No abnormalities detected
2000	3-1 Male	Killed day 14	No abnormalities detected
2000	3-2 Male	Killed day 14	No abnormalities detected
2000	3-3 Male	Killed day 14	No abnormalities detected
2000	2-0 Female	Killed day 14	No abnormalities detected
2000	4-0 Female	Killed day 14	No abnormalities detected
2000	4-1 Female	Killed day 14	No abnormalities detected
2000	4-2 Female	Killed day 14	No abnormalities detected
2000	4-3 Female	Killed day 14	No abnormalities detected

Individual Dermal reactions

Dose mg/kg	Animal & Sex	Observation	D1	D2	D3	D4	D5	D6	D7	D8	D9	D10	D11	D12	D13	D14
2000	1-0 Male	Erythema Oedema Other	0 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	3-0 Male	Erythema Oedema Other	0 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	3-1 Male	Erythema Oedema Other	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	3-2 Male	Erythema Oedema Other	0 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	3-3 Male	Erythema Oedema Other	0 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	2-0 Female	Erythema Oedema Other	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	4-0 Female	Erythema Oedema Other	1 0 0	1 0 0	1 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0  0 0	0 0 0	0 0 0	0 0 0
2000	4-1 Female	Erythema Oedema Other	2 0 0	2 0 0	2 0 Hd	1 0 Hd	0 0 Hd	0 0 Hd	0 0 Ss	0 0 Cf	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	4-2 Female	Erythema Oedema Other	1 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0
2000	4-3 Female	Erythema Oedema Other	1 0 0	1 0 0	1 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0	0 0 0

Where 0= No reactions, Hd = haemorrhage of dermal capillaries, Ss = Small superfical scabs, Cf = Crust formation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal median lethal dose (LD50) of the test material in the Wistar strain rat was found to be greater than 2,000 mg/kg

Executive summary:

An Acute dermal limit test (project # 3180/006) was commissioned by Nalco Ltd, UK and performed by Harlan Laboratories Ltd, UK in accorance with Method B4 of commision regulation 440/2008. The study was performed in the Wistar rat on PR-4758 technical grade (batch # FA0A0049). The study was performed in strict accordance with GLP. Standard protocols for animal husbandy and test method were followed and no deviations from the husbandy conditions or method recorded. Results indicate that the Acute dermal LD50 in the Wistar rat is > 2,000 mg/kg. No deaths were recorded, there were no clinical signs of systemic toxicity. Bodyweight gain was within normal parameters, no significant dermal irritation effects were recorded and no gross abnormalities noted at necrospy.