9-Octadecenoic acid (Z)-, sulfonated, potassium salts

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 26, 2014 to August 14, 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

9-Octadecenoic acid (Z)-, sulfonated, potassium salts

EC Number:

271-843-1

EC Name:

9-Octadecenoic acid (Z)-, sulfonated, potassium salts

Cas Number:

68609-93-8

Molecular formula:

A generic formula cannot be provided for this UVCB substance. The alkyl chain length of the sulfonated fatty acids range from C12-C22, however the major alkyl chain is C18.

IUPAC Name:

9-Octadecenoic acid (Z)-, sulfonated, potassium salts

Test material form:

other: powder/flakes with lumps

Details on test material:

- Name of test material (as cited in study report): 9-Octadecenoic acid (Z)-, sulfonated, potassium salts
- Physical state: Beige-yellow powder/flakes with lumps (i.e., determined at WIL Research Europe B.V.)
- Analytical purity: 100%
- Water content: 0.5%
- pH: 6 at concentration of 1g/L
- Lot/batch No.: 7495382
- Expiration date of the lot/batch: December 31, 2015
- Storage condition of test material: At room temperature in the dark
- Stability in vehicle: Water: Yes
- Solubility in vehicle: Water: Completely miscible

Test animals

Species:

rat

Strain:

other: Wistar strain, Crl:WI (Han) (outbred, SPF-Quality)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age: Young adult animals (approximately 10-12 weeks old) were selected. Females were nulliparous and non-pregnant
- Body weight: Body weight variation did not exceed +/- 20% of the sex mean
- Housing: Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet: Free access to pelleted rodent diet (i.e., SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 d. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm).
- Animal identification: Tail mark with indelible ink
- Health inspection: At least prior to dosing. It was ensured that the animals were healthy and that the skin to be treated was intact and free from any abnormality.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 to 24°C
- Humidity: 40 to 70%
- Air changes: 10 air changes/h
- Photoperiod: 12 h light/12 h dark cycle

IN-LIFE DATES: From June 26, 2014 to August 14, 2014

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

Treatment
- Method: Dermal application. Test substance (formulations) were stirred on a magnetic stirrer during application.

- Clipping: 1 d before exposure (i.e., Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.

- Application: The test substance formulation was applied in an area of approximately 10% of the total body surface, i.e. approximately 25 cm² for males and 18 cm² for females. The test substance formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D)*, successively covered with aluminum foil and Coban elastic bandage*. A piece of Micropore tape* was additionally used for fixation of the bandages in females only.
*. Manufacturers: Laboratoires Stella s.a., Liege, Belgium (surgical gauze) and 3M, St. Paul, Minnesota, U.S.A. (Coban & Micropore).

- Frequency: Single dosage, on Day 1.

- Dose level (volume): 200 mg/kg (10 mL/kg) bw
1,000 mg/kg (10 mL/kg) bw
2,000 mg/kg (10 mL/kg) bw

- Application period: 24 h, after which dressings were removed and the skin cleaned of residual test substance using tap water.

- Test substance preparation
Vehicle: Water (Elix, Millipore S.A.S., Molsheim, France)
Rationale: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.
Preparation: The formulations (w/w) were prepared within 4 h prior to dosing. Homogeneity was accomplished to a visually acceptable level. No correction was made for purity of the test substance.

Duration of exposure:

24 h

Doses:

200, 1,000 mg/kg and 2,000 mg/kg bw

No. of animals per sex per dose:

200 and 1,000 mg/kg bw: Three females/dose
2,000 mg/kg bw: Five/sex

Details on study design:

- Study design
Based on the test substance data available, it was considered that severe skin effects may occur after dermal application of the test substance. Therefore, the study was conducted in a step wise fashion in order to select the highest dose that could be tested without causing severe skin effects.
Initially, three females were dosed at 200 mg/kg bw. The absence or presence of systemic and local effects determined the next step. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups. The highest dose causing no severe effects was tested in 5 males and 5 females in total.

- Duration of observation period following administration: 14 d
- Frequency of observations:
- Mortality/Viability: Twice daily.

- Body weights: Days 1 (pre-administration), 8 and 15.

- Clinical signs: At periodic intervals on the day of dosing (i.e., Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).

- Necropsy: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

other: At 200 mg/kg, chromodacryorrhoea was noted for one animal on Day 1. At 1,000 mg/kg, general erythema was seen in the treated skin area of two animals on Days 2 and 3 and scales (on the flank of the animal) were noted for one animal on Days 7 and 8. At

Gross pathology:

At 200 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.
At 1,000 mg/kg, pelvic dilation and isolated gray-white focus/foci were found in the kidneys of one animal, at macroscopic post mortem examination.
At 2,000 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:

not classified

Conclusions:

Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats.

Executive summary:

A study was conducted to determine the acute dermal toxicity of the test substance in Wistar strain, Crl:WI (Han) rats according to OECD Guideline 402, in compliance with GLP. Based on available data, it was considered that severe skin effects may occur after dermal application of the test substance. Therefore, the study was conducted in a step wise fashion in order to select the highest dose that could be tested without causing severe skin effects. Initially, groups of three female rats received a single dermal dose of 200 or 1,000 mg/kg bw. Since no severe local or systemic effects were found at these treatment levels, a limit dose of 2,000 mg/kg bw was applied to skin of groups of five female and five male rats as a single dermal application for 24 hours. No mortality occurred and no effect on body weight gain was observed. No significant macroscopic abnormalities were seen at necropsy. However, chromodacryorrhoea was observed on Day 1 in one female and male and some local effects (general erythema and scales in the treated skin-area and/or on the flanks of the animals) were observed during the observation period (Van Huygevoort, 2014).

Under the study conditions, the dermal LD50 of the test substance was found to be >2,000 mg/kg bw in rats.