2,4,8,10-tetraoxa-3λ6,9λ6-dithiaspiro[5.5]undecane 3,3,9,9-tetraoxide

Endpoint summary

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

(17th December 2001)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Specific details on test material used for the study:

Test item: 2,4,8,10-Tetraoxa-3,9-dithiaspiro[5.5]undecane, 3,3,9,9-
tetraoxide
CAS: 201419-80-9
Lot number: AZ08AVL1
Active component: >99%
Appearance: crystalline solid, white
Expiration date: 21 September 2021
Storage conditions: room temperature, protected from humidity, well-closed
container

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Experimental Animals
Species and strain: Han:WIST rats
Source: TOXI COOP ZRT.
Cserkesz u. 90.
1103 Budapest, Hungary
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: The Wistar rats as a rodent is one of the standard species
of acute toxicity studies
Number of animals: 3 animals/group
Sex: Female, nulliparous and non pregnant animals
Age of animals: Young adult rat, 8 weeks old in first, second and third step
Body weight range
at starting (first step): 171 - 174 g
Body weight range
at starting (second step): 169 - 174 g
Body weight range
at starting (third step): 170 - 175 g
Acclimatization time: 5 days in first step, 6 days in second step and 7 days in
third step

Husbandry
Animal health: Only healthy animals were used for the study. Health
status was certified by the study director.
Room: 13/1
Housing: Group caging (3 animals/cage)
Cage type: Type III polypropylene/polycarbonate
Bedding: Certified laboratory bedding
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: above 10 air exchanges/hour by central air-condition system.
The temperature and relative humidity parameters were recorded daily during the study.

Route of administration:

oral: gavage

Vehicle:

other: sunflower oil

Remarks:

Name: Helianthi annui oleum raffinatum Batch number: 8006332001 Produced by: Magilab Kft. Date of expiration: 30.09.2021

Details on oral exposure:

Formulation
All doses were formulated in the vehicle. Concentration of formulations was adjusted to
maintain a treatment volume of 10 mL/kg bw. The test item was applied in a concentration
of 0.5 and 5 mg/mL. Formulations were prepared just before the administration and were
stirred continuously during the treatment.

Justification of the doses
Starting dose was selected on the basis of the available information about the test item. The
LD50 value is between 5 and 50 mg/kg bw according to the safety data sheet. The acute toxic
class method was carried out involving a stepwise procedure with the use of 5 mg/kg bw as
the starting dose in three female rats. No animal died in first step, so further three female rats
were treated with the same dose. No animal died in second step, too, so further three female
rats were treated with the higher dose (50 mg/kg bw). All animals died in third step, so the test
was finished, because the stopping criteria of Annex 2a of OECD Guideline No. 423
(presented in Appendix VII) was met.

Doses:

5 mg/kg bw
50 mg/kg bw

No. of animals per sex per dose:

3 female rats / dose

Control animals:

no

Details on study design:

Starting dose was selected on the basis of the available information about the test item. The
LD50 value is between 5 and 50 mg/kg bw according to the safety data sheet. The acute toxic
class method was carried out involving a stepwise procedure with the use of 5 mg/kg bw as
the starting dose in three female rats. No animal died in first step, so further three female rats
were treated with the same dose. No animal died in second step, too, so further three female
rats were treated with the higher dose (50 mg/kg bw). All animals died in third step, so the test
was finished, because the stopping criteria of Annex 2a of OECD Guideline No. 423
(presented in Appendix VII) was met.
Animals were weighed, observed for lethality and toxic symptoms for 14 days after the
treatment. Gross pathological examination was carried out in animals died on the treatment
day, as well as 15th day after the treatment in surviving animals.

Statistics:

The method used is not intended to allow the calculation of a precise LD50 value.

Preliminary study:

na

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

ca. 25 mg/kg bw

Based on:

test mat.

Mortality:

No lethality was noted at single oral dose of 5 mg/kg bw. ( 0/6 animals)
All female rats in step 3 (50 mg/kg bw) died on the treatment day 30 minutes after the treatment. (3/3 animals)

Clinical signs:

other: In group 1 treated with 5 mg/kg bw dose clinical sign of reaction comprised of decreased activity (6 cases of 57 observations), piloerection (6/57), tremor (4/57) and tonic convulsion (1/57). Decreased activity (score -1) and piloerection (score +1) occur

Gross pathology:

Six animals treated with 5 mg/kg bw dose survived until the scheduled necropsy on Day 15.
All of three rats treated with 50 mg/kg bw dose of the test item were necropsied as dead
animals on the treatment day.
External necropsy findings as froth around the mouth was detected in all animals of group 3
(50 mg/kg bw) and blood around the mouth was recorded in animal No.: 9712 of same
group. No changes were found related to the effect of the test item during the macroscopic
examination of 5 mg/kg bw dose.

Interpretation of results:

Category 2 based on GHS criteria

Remarks:

H300 CLP classification Acute tox category 2: H300

Conclusions:

CLP classification Acute tox category 2: H300 fatali if swallowed. The LD50 cut- off value is 25 mg/kg bw.
The method used is not intended to allow the calculation of a precise LD50 value.
The test item was ranked into classes of Globally Harmonized Classification System (GHS)
described in the OECD Guideline No. 423 as below:

Executive summary:

ACUTE ORAL TOXICITY STUDY(ACUTE TOXIC CLASS METHOD) OF TEST ITEM 2,4,8,10-TETRAOXA-3,9-DITHIASPIRO[5.5]UNDECANE, 3,3,9,9- TETRAOXIDE HAS BEEN PERFORMED IN WISTAR RATS ACCORDING TO OECD GUIDELINE 423.

Starting dose was selected on the basis of the available information about the test item. The
LD50value is between 5 and 50 mg/kg bw according to the safety data sheet. The acute toxic
class method was carried out involving a stepwise procedure with the use of 5 mg/kg bw as
the starting dose in three female rats. No animal died in first step, so further three female rats
were treated with the same dose. No animal died in second step, too, so further three female
rats were treated with the higher dose (50 mg/kg bw). All animals died in third step, so the test
was finished, because the stopping criteria of Annex 2a of OECD Guideline No. 423
(presented in Appendix VII) was met.
Animals were weighed, observed for lethality and toxic symptoms for 14 days after the
treatment. Gross pathological examination was carried out in animals died on the treatment
day, as well as 15th day after the treatment in surviving animals.
Lethality, Clinical symptoms and Body weight:
No lethality was noted at single oral dose of 5 mg/kg bw.
All female rats in step 3 (50 mg/kg bw) died on the treatment day 30 minutes after the
treatment.
In first step, CNS - and emotion symptoms (decreased activity, tremor, tonic convulsion)
and a disturbance of the autonomic functions (piloerection) were observed in animals on the
treatment day between 1 and 2 hours after the treatment.
In second step, CNS - and emotion symptoms (decreased activity, tremor, tonic convulsion,
clonic convulsion, closed eyes) and a disturbance of the autonomic functions (piloerection)
were observed in animals on the treatment day between 30 minutes and 2 hours after the
treatment.
In third step, CNS - and emotion symptoms (tonic- and clonic convulsion) and a disturbance
of the autonomic functions (dyspnoea) were observed in animals on the treatment day 30
minutes after the treatment. Besides, an indirect effect as frothy salivation was detected in
animals, which was consequence of convulsions.
The body weight development was undisturbed in all surviving animals.
Gross pathology:
All organs of the animals treated with 5 and 50 mg/kg bw proved to be free of treatment
related gross pathological changes, although it is necessary to note, that the external change
as frothy/bloody circumoral region was observed in dead animals.

Evaluation:
The method used is not intended to allow the calculation of a precise LD50value.
The test item was ranked into classes of Globally Harmonized Classification System (GHS)
described in the OECD Guideline No. 423 as below:
Table 1:

Dose (mg/kg bw)	Mortality (dead/treated)	LD50 (mg/kg bw)	GHS category
5	0/6	between 5 and 50	2
50	3/3

The LD50cut- off value is 25 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

25 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

(Q)SAR

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model, but not (completely) falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

QSAR borderline reliiable

Principles of method if other than guideline:

QSAR

GLP compliance:

no

Key result

Sex:

not specified

Dose descriptor:

LC50

Effect level:

ca. 1.36 mg/L air

Based on:

not specified

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Prediction Acute inhalation LC50 in rat equal to 1.36e+03 mg/m3/h (1.36 mg/L/h) It is classifed as acute toxicity category 4 inhalation: H332
Computational tool: Topkat
Acute inhalation toxicity on rat prediction was generated employing Topkat.
Topkat model for acute inhalation toxicity on rat provides LC50 predictions for rats by inhalation
administration route, based on PLS modelling methodology. For modelling purposes, critically reviewed and
selected experimental LC50 data, which were based on exposure times ranging from 0.5 to 14 hours, have
been normalized to per hour (assuming that within the range of adjustment, toxicity was proportional
to duration). The modelled endpoint is -log10(LC50) value. The unit of LC50 is mole/m3/h, which is
then converted to mg/m3/h. The prediction results of acute inhalation toxicity for 2,4,8,10-Tetraoxa-3,9-
dithiaspiro[5.5]undecane, 3,3,9,9-tetraoxide are illustrated in Table 5.15 and following discussed.

Executive summary:

Prediction Acute inhalation LC50 in rat equal to 1.36e+03 mg/m3/h (1.36 mg/L/h)

It is classifed as acute toxicity category 4 inhalation: H332

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

1 360 mg/m³ air

Quality of whole database:

Acute inhalation LC50 in rat equal to 1.36e+03 mg/m3/h (1.36 mg/L/h) Classified as CLP aucute toxicity inhalation category 4. H332

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)

Version / remarks:

9 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Specific details on test material used for the study:

Test item: 2,4,8,10-Tetraoxa-3,9-dithiaspiro[5.5]undecane, 3,3,9,9-tetraoxide
CAS: 201419-80-9
Lot number: AZ08AVL1
Active component: >99%
Appearance: crystalline solid, white
Expiration date: 21 September 2021
Storage conditions: room temperature, protected from humidity,
in well-closed container
pH (measured by
Toxi-Coop Zrt.): 5-6

Species:

rat

Strain:

Wistar

Remarks:

Han:WIST rats

Sex:

female

Details on test animals or test system and environmental conditions:

Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: above 10 air exchanges/hour by central air-condition system.
The temperature and relative humidity parameters were recorded daily during the study.

Type of coverage:

semiocclusive

Duration of exposure:

The test item was applied in undiluted form after pulverization and left in contact with the
skin for 24 hours, followed by a 14-day observation period.

Doses:

200 mg/kg bw , 1000 mg/kg bw , 2000 mg/kg bw

No. of animals per sex per dose:

1 female animal /200 mg/kg bw ,
3 female animals / 1000 mg/kg bw ,
1 female animals /2000 mg/kg bw

Details on study design:

At first, the range-finding study was performed. The starting dose was 200 mg/kg bw using
one animal. This animal did not die. The other doses were 1000 and 2000 mg/kg bw. The
animal of 1000 mg/kg bw did not die, too. The animal of 2000 mg/kg bw died. Therefore,
1000 mg/kg bw dose was used in main study

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

ca. 1 000 mg/kg bw

Based on:

test mat.

Mortality:

Inspection for signs of morbidity and mortality were made twice daily at the beginning and
end of the working day.
The initial dose was 200 mg/kg bw in the range-finding study. No mortality occurred after
the 24-hour dermal exposure to 2,4,8,10-Tetraoxa-3,9-dithiaspiro[5.5]undecane, 3,3,9,9-
tetraoxide in female rats treated with 200 and 1000 mg/kg bw dose of the range-finding
study. The animal treated with 2000 mg/kg bw dose died on the Day 1 in the range-finding
study.
The dose was 1000 mg/kg bw in main study. Both animals died. Animal No.: 1803 died on
Day 4 and animal No.: 1808 died on Day 2. The deaths seemed to be the consequence of
systemic toxic effects of the test item

Clinical signs:

convulsions

observations of tremors

Body weight:

lower than 10% body weight loss

Remarks:

The body weight of animal treated with 200 mg/kg bw corresponded to its species and age throughout the study. Body weight loss was observed in surviving animal (No.: 1801) of range-finding study between Day 0 and Day 7. Although, the body weight loss was below 10 % (approx. 6.1 %) and the body weight of this animal exceed the original body weight by the end of study, it can be evaluated as a toxic effect of the test item considering systemic clinical symptoms, too

Gross pathology:

One rat (No.: 1802) treated with 2000 mg/kg body weight of the test item spontaneously
died and were necropsied on Day 1. Two rats (No.: 1808, 1803) treated with 1000 mg/kg
body weight of the test item spontaneously died and were necropsied on Day 2 and on Day
4, respectively. One animal (No.: 1801) of the 1000 mg/kg body weight dose group and the
animal No.: 1800 dosed with 200 mg/kg body weight survived until the scheduled necropsy
on Day 15.
Internal necropsy findings were detected in range-finding study at 2000mg/kg bw and
external and internal changes were found in main study at 1000 mg/kg bw dose.
Range-finding study:
2000 mg/kg bw:
The colour of the lungs and liver were darker colour.
Main study:
1000 mg/kg bw:
Bloody and frothy discharge around the nose and mouth as external findings was observed
in animal No.: 1808. Attenuated mucous membrane in the stomach, haemorrhaged fundus,
bloody content in the intestines and autolysed organs were detected in both animals (No.:
1803, 1808). Granular cardia with erosion was recorded in animal No.: 1803. Intestines were
full of gas in animal No.: 1808 and marbled lungs was found in the same animal.

Other findings:

The animal treated with 2000 mg/kg bw died during the range-finding study. Two animals
treated with 1000 mg/kg bw died in the main study. No death occurred in the 200 mg/kg bw
dose.
The observed clinical signs in 1000 and 2000 mg/kg bw dose were related to the systemic
toxic effect of the test item.
There were no local clinical signs in all doses.
There was a change related to the toxic effect of the test item found in body weight and body
weight gain in surviving animal of 1000 mg/kg body weight dose on first week.
The pathological alterations observed in the non-surviving animals treated with 1000 mg/kg
bw or 2000 mg/kg bw were related to the effect of the test item, except autolysis. Autolysis
is a normal process after death. The external findings were consequence of indirect effect of
the test item, these were connected with convulsion.

Interpretation of results:

Category 3 based on GHS criteria

Remarks:

H311Toxic in contact with skin.

Conclusions:

Under the experimental conditions applied, the GHS category of the test item 2,4,8,10-
Tetraoxa-3,9-dithiaspiro[5.5]undecane, 3,3,9,9-tetraoxide is 3 in female Han:WIST rats
and the health hazard is H311: Toxic in contact with skin.

Executive summary:

An acute dermal toxicity study was performed with test item 2,4,8,10-Tetraoxa-3,9-
dithiaspiro[5.5]undecane, 3,3,9,9-tetraoxide in Han:WIST rats.
At first, the range-finding study was performed. The starting dose was 200 mg/kg bw using
one animal. This animal did not die. The other doses were 1000 and 2000 mg/kg bw. The
animal of 1000 mg/kg bw did not die, too. The animal of 2000 mg/kg bw died. Therefore,
1000 mg/kg bw dose was used in main study.
The test item was applied in undiluted form after pulverization and left in contact with the
skin for 24 hours, followed by a 14-day observation period.
Results of the main study:
Both animals died on Day 2 and on Day 4, respectively.
Systemic clinical symptoms as CNS (central nervous system) - and emotion symptoms
(decreased activity, tremor, clonic convulsion), disturbances of coordination (abnormal gait,
incoordination) and a disturbance of autonomic functions (piloerection) were observed
between Day 1 and Day 3.
The test item did not cause any dermal irritation symptom.
The body weight development could not be evaluated because of mortalities.
Autopsy revealed treatment related external alterations as bloody and frothy discharge
around the nose and mouth and internal alterations in the stomach (attenuated mucous
membrane, haemorrhage, granular cardia, erosion in the cardia), in the intestines (bloody
content and/or full of gas) and in the lungs (marbled) in died animals.
Under the experimental conditions applied, the GHS category of the test item 2,4,8,10-
Tetraoxa-3,9-dithiaspiro[5.5]undecane, 3,3,9,9-tetraoxide is 3 in female Han:WIST rats
and the health hazard is H311: Toxic in contact with skin.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

ca. 1 000 mg/kg bw

Quality of whole database:

1