Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 August 2015 to 01 September 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed in accordance with OECD, EU, US EPA & Japanese test guidelines in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
See Any other information
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
see Any other information
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
see Any other information
Qualifier:
according to
Guideline:
other: JMAFF Guidelines (2000), including the most recent revisions.
Deviations:
yes
Remarks:
see Any other information
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: pellets
Details on test material:
Identification: 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate]
Appearance: White to off white pellet
Batch: T3J07001
Purity/Composition: 97.8%
Test substance storage: At room temperature
Stable under storage conditions until: 11 September 2017 (retest date)
Chemical name (IUPAC): 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate]
Trade name: NAUGARD® 412S
CAS Number: 29598-76-3
Molecular formula: C65H124O8S4
Molecular weight: 1161.94
pH (1% in water, indicative range): 7.30-7.66 (determined by WIL Research Europe)

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species: Rat, Wistar strain Crl:WI (Han) (outbred, SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC).
Source: Charles River Deutschland, Sulzfeld, Germany.
Number of animals: 6 Females (nulliparous and non-pregnant). Each dose group consisted of 3 animals.
Age and body weight: Young adult animals (approx. 10-11 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification: Earmark and tail mark
Health inspection: At least prior to dosing. It was ensured that the animals were healthy and without any abnormality that might affect the study integrity.

Animal husbandry
Conditions
Environmental controls for the animal room are set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle: the photoperiod is between 07:00 and 19:00 hrs daily. The light/dark cycle may be interrupted for study related activities. Any variations to these conditions will be evaluated and maintained in the raw data.

Accommodation
Group housing of 3 animals per cage in labeled Makrolon cages (MIV 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).
Acclimatization period was at least 5 days before start of treatment under laboratory conditions.

Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
Water: Free access to tap water.

Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
Vehicle: Corn oil (Fagron Capelle a/d IJssel, The Netherlands) (specific gravity 0.92)
Rationale: The vehicle was selected based on trial preparations performed at WIL Research Europe and on test substance data supplied by the Sponsor.
There was no information available regarding the solubility or stability in vehicle.
Preparation: The preparations (w/w) were dosed within 4 hours after adding the vehicle to the test substance. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies.
Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test substance.

The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines.
The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.
Doses:
2000 mg/kg (10 mL/kg) body weight.
No. of animals per sex per dose:
3 animals
Control animals:
no
Details on study design:
Treatment
Method: Oral gavage, using plastic feeding tubes. The test item preparations were stirred on a magnetic stirrer during dosing.
Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.
Frequency: Single dosage on Day 1.
Dose level (volume): 2000 mg/kg (10 mL/kg) body weight.

Observations
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded:
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.
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Preliminary study:
Preiminary study not undertaken.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture (grade 1) and/or piloerection (grade 1) were noted for four females on Day 1 and/or 2.
Alopecia (grade 1), noted for one animal, from Day 6 onwards, is commonly seen in group housed rats and therefore no toxicological significance was attached to this finding.
Body weight:
The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
No further findings specified in the study report.

Any other information on results incl. tables

MORTALITY DATA

TEST DAY

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

HOURS AFTER TREATMENT

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

FEMALES 2000 MG/KG

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

FEMALES 2000 MG/KG

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

 

CLINICAL SIGNS

TEST DAY

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

HOURS AFTER TREATMENT

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MAX GRADE

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

FEMALES 2000 MG/KG

ANIMAL 1

No clinical signs noted

 

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

ANIMAL 2

Posture

-Hunched posture

Skin / fur

-Alopecia

 

 

(1)

 

(3)

 

 

1

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

 

 

-

 

1

ANIMAL 3

No clinical signs noted

 

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

 

-

FEMALES 2000 MG/KG

ANIMAL 4

Posture

-Hunched posture

Skin / fur

-Piloerection

 

 

(1)

 

(1)

 

 

1

 

-

 

 

1

 

1

 

 

1

 

1

 

 

1

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

ANIMAL 5

Posture

-Hunched Posture

Skin / fur

-Piloerection

 

 

(1)

 

(1)

 

 

1

 

-

 

 

1

 

1

 

 

1

 

1

 

 

1

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

ANIMAL 6

Posture

-Hunched Posture

Skin / fur

-Piloerection

 

 

(1)

 

(1)

 

 

1

 

-

 

 

1

 

1

 

 

1

 

1

 

 

1

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

 

 

-

 

-

- = Sign not observed

 

BODY WEIGHTS (GRAM)

SEX/DOSE LEVEL

ANIMAL

DAY 1

DAY 8

DAY 15

FEMALES 2000 MG/KG

 

1

2

3

 

MEAN

ST. DEV.

N

 

196

189

191

 

192

4

3

 

215

214

229

 

219

8

3

 

218

218

242

 

226

14

3

FEMALES 2000 MG/KG

 

4

5

6

 

MEAN

ST. DEV.

N

 

177

156

183

 

172

14

3

 

201

173

208

 

194

19

3

 

215

187

218

 

207

17

3

 

MACROSCOPIC FINDINGS

ANIMAL ORGAN

FINDING

DAY OF DEATH

FEMALES 2000 MG/KG

1

No findings noted

Scheduled necropsy Day 15 after treatment

2

No findings noted

Scheduled necropsy Day 15 after treatment

3

No findings noted

Scheduled necropsy Day 15 after treatment

FEMALES 2000 MG/KG

4

No findings noted

Scheduled necropsy Day 15 after treatment

5

No findings noted

Scheduled necropsy Day 15 after treatment

6

No findings noted

Scheduled necropsy Day 15 after treatment

 

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 value of 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] in Wistar rats was established to exceed 2000 mg/kg body weight.
Executive summary:

Assessment of acute oral toxicity with 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] in the rat (Acute Toxic Class Method).

 

The study was carried out based on the guidelines described in:

OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method"

Commission Regulation (EC) No 440/2008, B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method"

EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

JMAFF Guidelines (2000), including the most recent revisions.

 

2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight.

Macroscopic examination was performed after terminal sacrifice (Day 15).

 

No mortality occurred.

Hunched posture (grade 1) and/or piloerection (grade 1) were noted for four females on Day 1 and/or 2.

The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

No abnormalities were found at macroscopic post mortem examination of the animals.

 

The oral LD50 value of 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] in Wistar rats was established to exceed 2000 mg/kg body weight.

 

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

 

Based on these results, 2,2-Bis[[3-(dodecylthio)-1-oxopropoxy]methyl]propane-1,3-diyl bis[3-(dodecylthio)propionate] does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).