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Description of key information

Oral:

In an acute oral toxicicty study conducted according to OECD Guideline 423 in rats, an oral LD50 > 2000 mg/kg bw was determined (UN GHS: no classification) (reference 7.2.1-1).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 July 2017 - 05 September 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
Crl:WI rats
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90., Hungary
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats, 11 weeks old
- Weight at study initiation: 236 - 247 g (first group), 235 - 242 g (second group)
- Fasting period before study: The day before treatment the animals were fasted. The food but not water was withheld overnight.
- Housing: 3 animals/sex/cage in Type III polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Diet: ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum.
- Water: tap water from watering bottles ad libitum.
- Acclimation period: 26 days for group 1 and 27 days for group 2

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): > 10
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
other: Helianthi annui oleum raffinatum
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle: 10 mL/kg bw
- Lot/batch no.: 1607-4569

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: All doses were formulated in the vehicle. Concentration of formulations were adjusted to maintain a treatment volume of 10 mL/kg bw. Formulations were prepared just before the administration and stirred continuously during the treatment.

CLASS METHOD
- Rationale for the selection of the starting dose: The starting dose was selected on the basis of the available information on the test item.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Inspection for signs of morbidity and mortality were made twice daily at the beginning and end of the working day. Animals were observed individually after dosing once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and once per day for 14 days thereafter. The body weight was recorded on day 0 (shortly before the treatment), on day 7 and on day 15 with a precision of 1 g, respectively.
- Necropsy of survivors performed: yes, all animals were exsanguinated under isoflurane anaesthesia. After examination of the external appearance the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross pathological changes were recorded for each animal on the post mortem record sheets.
- Other examinations performed: clinical signs, body weight, gross pathology
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The test item did not induce mortality following a single oral administration to female rats at a dose of 2000 mg/kg bw (group 1 and 2, step 1 and 2). All rats in step 1 and step 2 survived until the end of the 14-day observation period.
Clinical signs:
In group 1 treated with 2000 mg/kg bw, clinical signs comprised of abnormal gait (5 cases out of 57 observations), incoordination (5/57), decreased activity (3/57), prone position (3/57), limb position (3/57), paralysis (2/57), decreased righting reflex (3/57), decreased grip- and limb tone (3/57), decreased body tone (3/57), decreased abdominal tone (3/57), closed eyes (1/57), piloerection (1/57) and dyspnoea (2/57). Abnormal gait (score +2, +3, +4) and incoordination (score +2, +3, +4) were observed in all animals. Prone position (score +3, +2), limb position (score +2, +3, +4), incoordination (score +4, +3), paralysis (score +2, +3), decreased righting reflex (score -1, -2, -3), decreased grip- and limb tone (score -2, -3), decreased body tone (score -2, -3), decreased abdominal tone (score -2, -3), closed eyes (score +2), piloerection (score +2) and dyspnoea (score +2, +1) were found in animal No.: 6442. These symptoms were observed on the treatment day between 2 and 4 hours after the treatment and they were connected with the test item toxic effect.
In group 2 treated with 2000 mg/kg bw clinical signs comprised of abnormal gait (18 cases out of 57 observations), incoordination (18/57), decreased activity (18/57), prone position (7/57), limb position (8/57), decreased righting reflex (8/57), decreased grip- and limb tone (13/57), decreased body tone (13/57) and decreased abdominal tone (13/57). These symptoms were observed in all animals. The scores were as follows: decreased activity (-1, -2), abnormal gait (+1, +2, +3), prone position (+1, +2, +3), limb position (score +1, +2, +3), incoordination (+1, +2, +3), decreased righting reflex (-1, -2, -3), decreased grip- and limb tone (-1, -2, -3), decreased body tone (-1, -2) and decreased abdominal tone (-1, -2). These symptoms were observed between the treatment day and day 1 and they were connected with the test item toxic effect.
Body weight:
In group 1 and 2 (2000 mg/kg bw) the mean body weight and body weight gain of the animals corresponded to their species and age throughout the study.
Gross pathology:
No pathological changes were found related to the test item during the macroscopic examination of animals. Two animals showed hydrometra, which is a physiological finding and connected to the cycle of the animal.
Interpretation of results:
GHS criteria not met
Conclusions:
In an acute oral toxicicty study conducted according to OECD Guideline 423 in rats, an oral LD50 > 2000 mg/kg bw was determined.
Executive summary:

In an acute oral toxicity study conducted according to OECD Guideline 423 (class method), two groups of three female Crl:WI rats each were sequentially treated via oral gavage with 2000 mg/kg bw 2,4-dithiapentane. No lethality was noted in any test group following administration. CNS- and emotion symptoms (decreased activity, paralysis, closed eyes), decreased reflex- and muscular tension (righting reflex, grip- and limb tone, body tone, abdominal tone), disturbances of the coordination (prone position, incoordination, abnormal gait, limb position) and disturbances of the autonomic functions (piloerection, dyspnoea) were observed in animals of the first group on the treatment day between 2 and 4 hours after the treatment. In the second group, CNS- and emotion symptom (decreased activity), decreased reflex- and muscular tension (righting reflex, grip- and limb tone, body tone, abdominal tone) and disturbances of the coordination (prone position, incoordination, abnormal gait, limb position) were observed in animals between the treatment day and day 1. No pathological changes were observed after gross pathology of the animals.

The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study with the test item in rats the determined LD50 is above 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
Klimisch code 1

Additional information

In an acute oral toxicity study conducted according to OECD Guideline 423 (class method), two groups of three female Crl:WI rats each were sequentially treated via oral gavage with 2000 mg/kg bw of the test item (reference 7.2.1-1). No lethality was noted in any test group following administration. CNS- and emotion symptoms (decreased activity, paralysis, closed eyes), decreased reflex- and muscular tension (righting reflex, grip- and limb tone, body tone, abdominal tone), disturbances of the coordination (prone position, incoordination, abnormal gait, limb position) and disturbances of the autonomic functions (piloerection, dyspnoea) were observed in animals of the first group on the treatment day between 2 and 4 hours after the treatment. In the second group, CNS- and emotion symptom (decreased activity), decreased reflex- and muscular tension (righting reflex, grip- and limb tone, body tone, abdominal tone) and disturbances of the coordination (prone position, incoordination, abnormal gait, limb position) were observed in animals between the treatment day and day 1. No pathological changes were observed after gross pathology of the animals.

The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study in rats the determined LD50 of the test item is above 2000 mg/kg bw (UN GHS: no classification).

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.

As a result the substance is not considered to be classified for acute oral toxicity under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.