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Toxicological information

Acute Toxicity: dermal

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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 June 2016 to 05 July 2016
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
clinical observations conducted at 2 and 5 hours after application with no impact on results or integrity of the study
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
L 142
Deviations:
yes
Remarks:
clinical observations conducted at 2 and 5 hours after application with no impact on results or integrity of the study
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
EPA 712-C-98-192, August 1998
Deviations:
yes
Remarks:
clinical observations conducted at 2 and 5 hours after application with no impact on results or integrity of the study
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Appearance/physical state: Amber liquid
- Storage conditions: Controlled room temperature (15-25 ºC, below 70 RH%)

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
EXPERIMENTAL ANIMALS
- Species and strain: Sprague Dawley Rat (Crl:CD(SD))
- Source: Charles River Laboratories, Research Models and Services GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany.
- Sex: Male and female, female rats were nulliparous and non-pregnant.
- Age of animals at study start: Young adult rats
- Body weight range at dosing: 215 to 247 g
- Acclimatisation period: 5 days

HUSBANDRY
- Animal health: Only healthy animals were used for the study. The veterinarian certified the health status.
- Room-Box: 245/9
- Housing: Individual caging
- Cage type: Type II. polypropylene/polycarbonate
- Bedding: Lignocel ¾S Hygienic Animal Bedding (produced by J. Rettenmaier & Söhne GmbH + Co.KG, Germany) was available to animals during the study. A copy of the Certificate of Analysis is retained in the archives at CiToxLAB Hungary Ltd.
- Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
- Temperature: 20.7 to 24.6 °C
- Relative humidity: 39 to 70 %
- Temperature and relative humidity were recorded twice daily during the study.
- Ventilation: 15 to 20 air exchanges/hour
- Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.

ANIMAL IDENTIFICATION
- The individual identification was performed using numbers written on the tail with a marker pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.' s Master File for each animal allocated to the treatment groups.
- Cages were identified by cards containing information about study code, sex, dose group, cage number and individual animal number.

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST ITEM ADMINISTRATION
- The test item was not expected to be lethal at 2000 mg/kg bw and a limit test was therefore performed.
- A single dermal application was made and was followed by a fourteen-day observation period (male animals cages 1 to 5; female animals cages 6 to 10).

PROCEDURE
- The back of each animal was shaved (approximately 10 % area of the total body surface) approximately 24 hours prior to treatment.
- Test item was applied as a single dose to the shaved skin and remained in contact with the skin for the 24-hour exposure period.
- Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.
- At the end of the exposure period, the area of skin treated with the test item was washed with water of body temperature.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
FOOD AND WATER SUPPLY
- Animals received ssniff SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany (Batch No.: 278 5652, expiry date: 30 November 2016), ad libitum, and tap water from the municipal supply, as for human consumption from a 500 ml bottles, ad libitum. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study and the water was considered fit for human consumption.
The supplier provided an analytical certificate for the batch used and copy certificates were archived with the raw data.
- Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A. u. 36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

CLINICAL OBSERVATIONS
- Clinical observations were performed on the day of treatment at 2 and 5 hours after application of the test item and once each day for 14 days thereafter.
- Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

SKIN IRRITATION
- Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.

MEASUREMENT OF BODY WEIGHT
- Body weights were recorded on Day 0 (before test item administration) and on Days 7 and 14 just before necropsy.

NECROPSY
- Macroscopic examination was performed on all animals. All animals were anaesthetised with pentobarbital sodium and exsanguinated.
- After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.
Statistics:
EVALUATION OF DATA
- The method used was not intended to allow the calculation of a precise Ld50 value.
- Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.

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:
- The test item did not cause mortality at the dose level of 2000 mg/kg bw.
Clinical signs:
- There were no systemic clinical signs noted in any animal throughout the study (see Table 1, attached).
Body weight:
- There were no treatment related effects on body weight or body weight gain during the observation period (see Table 2, attached).
Gross pathology:
- No macroscopic observations were noted at a dose level of 2000 mg/kg bw (see Table 3 and Appendix 1, attached).
Other findings:
LOCAL DERMAL EFFECTS
- Treatment with test item at the dose level of 2000 mg/kg bw caused oedema (oedema score: 1 or 2) (10/10), erythema (erythema score: 1 or 2) (8/10) and crust (1/10) in rats (see Table 1, attached).
- All rats were symptom free from Day 2 (3/10) or Day 3 (3/10) or Day 5 (4/10).

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item was found to be > 2000 mg/kg body weight in male and female Sprague Dawley rats.
Executive summary:

GUIDELINE

An acute dermal toxicity study was performed with test item in Sprague Dawley rats, in compliance with OECD Guideline No 402, EU Method B.3 and OPPTS 870.1200.

 

METHODS

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as a single dermal 24-hour exposure followed by a 14-day observation period. Clinical observations were performed on all animals at 2 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

 

RESULTS

The test item did not cause mortality at the dose level of 2000 mg/kg bw and no systemic clinical signs noted in any animal throughout the study. Treatment at a dose level of 2000 mg/kg bw caused oedema (oedema score: 1 or 2) (10/10), erythema (erythema score: 1 or 2) (8/10) and crust (1/10) in rats. All rats were symptom free from Day 2 (3/10) or Day 3 (3/10) or Day 5 (4/10). There were no treatment related effects on body weight or body weight gain during the observation period and no macroscopic observations were noted at a dose level of 2000 mg/kg bw.

 

CONCLUSION

The acute dermal median lethal dose (LD50) of the test item was found to be > 2000 mg/kg body weight in male and female Sprague Dawley rats.