Amines, di-C16-18-alkyl

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From the 23-Sep-2009 and 13-Oct-2009.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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:

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-(C16-C18)alkyl(C16-C18)alkane-1-amine

EC Number:

629-721-4

Cas Number:

308062-60-4

Molecular formula:

No molecular formula

IUPAC Name:

N-(C16-C18)alkyl(C16-C18)alkane-1-amine

Test material form:

solid

Details on test material:

Chemical name: Amines, di-C16-18 (even numbered) alkyl
EC no.: 629-721-4

To the best of knowledge, the sample used is representative to the boundary composition shared and agreed by each registrant.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test System
Animals: Rat, RccHan: WIST(SPF)

Rationale: Recognized by international guidelines as a recommended test system.

Breeder: Harlan Laboratories B.V. Kreuzelweg 53 5961 NM Horst / The Netherlands Postbus 6174 5960 AD Horst / The Netherlands

Number of Animals per Group: 5 males and 5 females

Total Number of Animals: 5 males and 5 females

Age (when treated): Males: 9 weeks Females: 11 weeks

Body Weight Range (when treated): 246.3 g – 256.0 g (males) 194.5 g – 201.2 g (females)

Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked at acclimatization start.

Randomization: Selected by hand at time of delivery. No computer generated randomization program.

Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
Allocation

The animals were distributed as follows:

Dose Level Sex Group Animal Number
2000 mg/kg M 1 1-5
2000 mg/kg F 2 6-10



Husbandry

Room Number: 0105 / Harlan Laboratories Ltd., Füllinsdorf

Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 22 ± 3 °C and for relative humidity between 30-70% (values above 70% during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.

Accommodation: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding (‘Lignocel’ J. Rettenmaier&Söhne GmbH&CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) during treatment and observation.

Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 30/09 (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) ad libitum.
Results of analyses for contaminants are archived at Harlan Laboratories Ltd.

Water: Community tap water from Füllinsdorf ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at Harlan Laboratories Ltd.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

corn oil

Details on dermal exposure:

One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10% of the total body surface.

Only those animals without injury or irritation on the skin were used in the test.

Vehicle
The following information was provided by Harlan Laboratories Ltd.:

Identification: Corn oil

Description: Yellowish oily liquid

Batch Number: 049103168

Source: Carl Roth GmbH & Co. 76185 Karlsruhe / Germany

Stability of the Vehicle: Stable under storage conditions

Expiry Date: 31-Jan-2014

Storage Conditions: At room temperature (range of 20 ± 5 °C), light protected.

Safety Precautions: Routine hygienic procedures were used to ensure the

health and safety of the personnel.
The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. This formulation trial is excluded from the statement of compliance.

Preparation of Dose Formulations
Dose levels are in terms of the test item as supplied by the Sponsor. The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume). The formulation was prepared shortly before the application using a magnetic stirrer, a spatula and an Ultra-Turrax as homogenizers. Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a surgical gauze pad (ca. 5 x 5 cm) held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and an elastic adhesive restrainer bandage wrapped around the abdomen.

The application volume was 6 mL/kg.

After the 24-hour application period, the dressing was removed and the skin was flushed with lukewarm water and drapped off with disposable paper towels. The skin reaction was assessed.

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

PURPOSE
The purpose of this study was to assess the acute dermal toxicity of Amines, bis(hydrogenated tallow alkyl) (CAS number: 61789-79-5) when administered to rats by a single semi-occlusive dermal application, followed by an observation period of 14 days. This study should provide a rational basis for risk assessment.

Rationale
Dermal administration was used as this is one possible route of human exposure during manufacture, handling and use of the test item.

Five male and five female RccHan:WIST (SPF) rats were treated with Amines, bis(hydrogenated tallow alkyl) (CAS number: 61789-79-5) at 2000 mg/kg by dermal application. The test item was formulated in corn oil at a concentration of 0.33 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Local signs were noted once daily from test day 2 to 15. Mortality/viability was recorded within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

Necropsy
All animals were killed at the end of the observation period by carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained.

Determination and Scoring of Clinical Signs and Local Dermal Signs
Observation
Data were summarized in tabular form, showing for each individual animal the clinical and local signs at each measurement interval. All findings were described.
The skin reaction was assessed according to the numerical scoring system listed in the Commission Regulation (EC) No 440/2008 B.4

Data Compilation
Body weights were recorded on-line.

Mortality/viability, clinical and local dermal signs were recorded on data sheets.

Macroscopic findings were compiled into the RCC Tox Computer System during recording.

The RCC Tox Computer System (RCC-Tox-Lims) had been validated with respect to data collection, storage and retrievability.

Statistics:

No statistical analysis was performed.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: 95 % CL not reported

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No clinical signs were observed during the course of the study. Well-defined erythema at the application sites was observed in all animals at day 2 (just after removal of the 24-hour application dressing) and lasted as well-defined to very slight up to

Gross pathology:

No macroscopic findings were recorded at necropsy.

Any other information on results incl. tables

Result tables are attached to this report.

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

The median lethal dose of Amines, di-C16-18 (even-numbered) alkyl (CAS No. 308062-60-4) after single dermal administration to rats of both sexes, observed over a period of 14 days, is:
LD50 (rat): greater than 2000 mg/kg body weight
Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, Amines, di-C16-18 (even-numbered) alkyl (CAS No. 308062-60-4) does not have to be classified and labelled with respect to acute dermal toxicity in the rat.

Executive summary:

Five male and five female RccHan:WIST (SPF) rats were treated with Amines, di-C16-18 (even-numbered) alkyl (CAS No. 308062-60-4) at 2000 mg/kg by dermal application. The test item was formulated in corn oil at a concentration of 0.33 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Local signs were noted once daily from test day 2 to 15. Mortality/viability was recorded within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study.

No clinical signs were observed during the course of the study.

Different local dermal signs were observed during the course of the study, including well defined to very slight erythema, scaling, scabs and fissures at the application sites. The test item did not cause any staining of the treated skin.

The body weight of the animals was within the range commonly recorded for this strain and age.

No macroscopic findings were observed at necropsy.

The median lethal dose of Amines, di-C16-18 (even-numbered) alkyl (CAS No. 308062-60-4) after single dermal administration to rats of both sexes, observed over a period of 14 days, is: LD50 (rat): greater than 2000 mg/kg body weight Based upon the classification criteria according to the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, Amines, di-C16-18 (even-numbered) alkyl (CAS No. 308062-60-4) does not have to be classified and labelled with respect to acute dermal toxicity in the rat.