Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 April 2017 to 25 April 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
OECD GUIDELINES FOR TESTING OF CHEMICALS (No.: 402, 24th Feb. 1987)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
Commission Regulation (EC) No 440/2008, B.3 (L 142, 30 May 2008)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
OPPTS 870.1200 (EPA 712-C-98-192, August 1998)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
Name: 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate]
CAS number: 29598-76-3
Batch/Lot Number: 1005151109
Description: White to off-white pellet
Manufacture date: 13 November 2015
Expiry date: 12 November 2019
Purity*: Considered as 100%
Storage condition: Controlled room temperature (15-25 ºC, below 70 RH%)
Safety precautions: Routine safety precautions (lab coat, gloves, safety glasses, face mask) for unknown materials were applied to assure personnel health and safety.
*No correction for purity of the test item was applied.
Specific details on test material used for the study:
No further details specified.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Species and strain: Crl:WI Wistar rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld, Germany
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Justification of strain: The Wistar rat is one of the standard rodent species used in acute toxicity studies.
Number of animals: 5 animals/sex
Sex: Male and female, female rats were nulliparous and non-pregnant.
Age of animals at dosing: Young adult rats
Body weight range at dosing: Between 218 g and 254 g
Acclimation time: 5 days

Husbandry
Animal health: Only healthy animals were used for the study.
The staff Veterinarian certified the health status.
Room-Box: 242/7
Housing: Individual caging
Cage type: Type II. polypropylene/polycarbonate
Bedding: Lignocel 3/4-S Hygienic Animal Bedding was available to animals during the study. Copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Nesting: Nest building material Arbocel Crinklets natural was available to animals during the study. Copy of the Certificate of Analysis is retained in the archive at CiToxLAB Hungary Ltd.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 19.4–24.7°C
Relative humidity: 30–58%
Ventilation: 15-20 air exchanges/hour
Enrichment: Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities.
Temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply
Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance", ad libitum, and tap water from the municipal supply, as for human consumption from a 500 mL bottle, ad libitum. The food is 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 batch of feed employed in the study was as follow:
• 285 17890, expiry date: 31 August 2017
The supplier provided an analytical certificate for the batch used. A copy of the certificate will be 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. The quality control results are retained in the archives at CiToxLAB Hungary Ltd.

Animal Identification
Animals were individually identified using numbers written on the tail with an indelible pen. The numbers were given on the basis of CiToxLAB Hungary Ltd.'s Master File for each animal allocated to the treatment groups. The 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:
water
Details on dermal exposure:
Formulation
The test item was powdered and administered as a single dose. Sufficient water was used to dampen the test material to ensure good contact with the skin.

Procedure
The back of each animal was shaved (approximately 10% area of the total body surface) approximately 24 hours prior to treatment. The test item was applied to the shaved skin as a single dose 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 treated area of skin with the test item was washed with water at body temperature.
Duration of exposure:
A single dermal application was made.
Doses:
The test item was not expected to be lethal at 2000 mg/kg bw. A limit test was therefore performed.
No. of animals per sex per dose:
10 (5 males/5 females)
Control animals:
not required
Details on study design:
OBSERVATIONS
Clinical Observations
Clinical observations were performed on the day of treatment at 1 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
The 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.
Following confirmation of death, 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:
The method used was not intended to allow the calculation of a precise LD50 value.
Body weight and body weight gain are summarised in tabular form. Clinical signs and necropsy findings are described and summarised in tabular form.

Results and discussion

Effect levels
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.
Body weight:
There were no treatment related effects on body weight or body weight gain during the observation period.
Gross pathology:
There was no evidence of any gross observations at a dose level of 2000 mg/kg bw.
Other findings:
Local Dermal Signs
No adverse local dermal signs were observed after treatment with the test item or during the 14 days observation period.

Any other information on results incl. tables

Clinical Observations

DOSE LEVEL: 2000 mg/kg bw                                                                                          SEX: MALE

Cage No.

Animal No.

Observations

Observations days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

1

7960

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

2

7961

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

3

7962

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

4

7963

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

5

7964

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

DOSE LEVEL: 2000 mg/kg bw                                                                                            SEX: FEMALE

Cage No.

Animal No.

Observations

Observations days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

6

7975

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

7

7976

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

8

7977

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

9

7978

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

10

7979

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

Remarks:   + = present

h = hour                  Treatment day = Day 0

Frequency of observation = number of occurrence of observation / total number of observations

 

Body Weight Data

DOSE LEVEL: 200 mg/kg bw                                     SEX: MALE

Cage No.

Animal No.

Body weight (g)

Days

Body Weight Gain (g)

0

7

14

0-7

7-14

0-14

1

7960

220

252

290

32

38

70

2

7961

244

300

353

56

53

109

3

7962

251

300

350

49

50

99

4

7963

230

279

325

49

46

95

5

7964

224

263

296

39

33

72

Mean:

233.8

278.8

322.8

45.0

44.0

89.0

Standard deviation:

13.2

21.6

29.4

9.5

8.3

17.2

DOSE LEVEL: 200 mg/kg bw                                   SEX: FEMALE

Cage No.

Animal No.

Body weight (g)

Days

Body Weight Gain (g)

0

7

14

0-7

7-14

0-14

6

7975

233

235

247

2

12

14

7

7976

222

227

234

5

7

12

8

7977

218

235

252

17

17

34

9

7978

232

233

239

1

6

7

10

7979

254

259

302

5

43

48

Mean:

231.8

237.8

254.8

6.0

17.0

23.0

Standard deviation:

14.0

12.3

27.3

6.4

15.2

17.3

 

Macroscopic Findings

DOSE LEVEL: 2000 mg/kg bw                                                                             SEX: MALE

Cage No.

Animal No.

Necropsy Date/ Necropsy Day

External Observations

Internal Observations

Organ / Tissue

1

7960

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

2

7961

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

3

7962

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

4

7963

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

5

7964

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

DOSE LEVEL: 2000 mg/kg bw                                                                                    SEX: FEMALE

Cage No.

Animal No.

Necropsy Date/ Necropsy Day

External Observations

Internal Observations

Organ / Tissue

6

7975

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

7

7976

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

8

7977

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

9

7978

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

10

7979

25 April 2017

Day 14

No external observations

No internal observations

Not applicable

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test item 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] was found to be greater than 2000 mg/kg body weight in male and female Crl:WI rats.
Executive summary:

An acute dermal toxicity study was performed with the test item 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] in Crl:WI Wistar rats, in compliance with OECDGuideline No.: 402.

 

A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was powdered and applied as a single dermal 24-hour exposure followed by a 14-day observation period.

 

Clinical observations were performed on all animals at 1 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 (before necropsy). Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

 

The results of the study were summarised as follows:

Mortality

Test item did not cause mortality at the dose level of 2000 mg/kg bw.

 

Systemic clinical signs

There were no systemic clinical signs noted in any animal throughout the study.

 

Local dermal signs

No adverse local dermal signs were observed after treatment with the test item or during the 14 days observation period.

 

Body weight and body weight gain

There were no treatment related effects on body weight or body weight gain during the observation period.

 

Necropsy

There was no evidence of any gross observations at a dose level of 2000 mg/kg bw.

 

Conclusions

The acute dermal median lethal dose (LD50) of the test item 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] was found to be greater than 2000 mg/kg body weight in male and female Crl:WI rats.

 

According to the GHS criteria, 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] can be ranked as "Category 5" or "Unclassified" for acute dermal exposure.