Phenol, 2,4-dinitro-, sulfurized, thiosulfonated

Endpoint summary

Description of key information

In an in vivo acute oral toxicity assay (acute toxicity class method, ATC) according to OECD guideline 423, an LD50 of above 2000 mg/kg bw was determined.

In accordance with Regulation (EG) No 1907/2006, Annex VIII, column 2 a dermal or inhalation toxicity study was not performed.

However, considering the weight of evidence, LD50/LC50 values above 2000 mg/kg bw or 5 mg/L can be expected.

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 May - 05 July, 2017

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:

17 December 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

30 May 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Expiration date: 30.11.2021

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI rats (SPF)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: TOXI COOP ZRT. Cserkesz u. 90., 1103 Budapest, Hungary
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rat, 8 weeks old in first and second step
- Weight at study initiation: first step 176 - 180 g; second step 200 - 202 g
- Fasting period before study: the day before administration
- Housing: Group caging (3 animals/cage) in Type II cages.
- Diet: ssniff® SM R/M-Z+H complete diet for rats and mice; ssniff Spezialdiäten GmbH, 59494 Soest, Germany, ad libitum
- Water: tap water from municipal supply, as for human consumption from bottle, ad libitum.
- Acclimation period: 6 days in the first step and 7 days in the second step
- Animal health: Only healthy animals were used for the study. Health status was certified by the study director.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes: above 10 air exchanges/hour by central air-condition system.
- Photoperiod (hrs dark / hrs light): 12/12, from 6.00 a.m. to 6.00 p.m.

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua purificata

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle: treatment volume of 10 mL/kg bw
- Batch no.: 1703-5503

CLASS METHOD
- Rationale for the selection of the starting dose: Starting dose was selected on the basis of the available information about the test item.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 females (3 per group)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed individually after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and twice each day for 14 days thereafter. The body weights were recorded on day 0 (just before the treatment), on day 7 and on day 15.
- Necropsy of survivors performed: yes. At the end of the observation period all survivor rats were sacrificed under isofluran anaesthesia. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed, and any abnormality was recorded with details of its location, colour, shape and size.
- Other examinations performed: Individual observations were performed on the skin and fur, eyes and mucous membranes and also 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.

Statistics:

The mean of the body weight and body weight gain were calculated by Excel spreadsheet software.
The method used is not intended to allow statistical evaluation and the calculation of a precise LD50 value.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 2 000 - <= 5 000 mg/kg bw

Based on:

act. ingr.

Remarks on result:

other: GHS category: 5 or unclassified

Mortality:

No death occurred at 2000 mg/kg bw single oral dose. All female rats in step 1 and step 2 survived until the end of the 14-day observation period.

Clinical signs:

other: In group 1 treated with 2000 mg/kg bw dose clinical sign of reaction comprised black faeces (3 cases of 57 observations) and black colored diarrhoea (8/57). Black faeces (score +1) were observed in all animals on Day 1. Black colored diarrhoea (score +1,

Gross pathology:

All animals treated with 2000 mg/kg bw survived until the scheduled necropsy on Day 15.
At necropsy pale liver was observed in one animal. This alteration was not considered to be related to the test item, but was regarded as an individual variation. Slight hydrometra was found in one animal, which is a physiological finding and related to the estrous cycle of the animal. No pathological changes were found related to the treatment with the test item during the macroscopic examination of animals treated with 2000 mg/kg bw.

Table 2: Summary of Mortality, Post-treatment observation period (14 days) 

Groups	Treatment		Lethality
	Test Item	Dose (mg/kg bw)	Females
1	Solubilised Sulphur Black 1 Step 1	2000	0/3
2	Solubilised Sulphur Black 1 Step 2	2000	0/3

Table 3: Summary of Clinical Symptoms

Groups	Treatment		Symptoms	Incidence
	Test Item	Dose (mg/kg bw)
1	Solubilised Sulphur Black 1 Step 1	2000	Black faeces	3/57
			Diarrhoea (black)	8/57
			Normal	46/57
2	Solubilised Sulphur Black 1 Step 2	2000	Black faeces	3/57
			Diarrhoea (black)	4/57
			Normal	50/57

 

Remark:        Incidence = Number of symptoms/Summarized number of observations inside the group

                         Summarized number of observations inside the group = (number of observations of first animal) + (number of observations of second animal) + (number of observations of third animal)

Table 4: Summary of Body Weights (g)

 

FEMALES		Day 0	Day 7	Day 15
Group 1: Solubilised Sulphur Black 1
2000 mg/kg bw, Step 1
Group size:		3	3	3
Mean: (g)		177.7	205.0	222.0
SD:		2.08	7.94	2.00
FEMALES		Day 0	Day 7	Day 15
Group 2:  Solubilised Sulphur Black 1
2000 mg/kg bw, Step 2
Group size:		3	3	3
Mean: (g)		201.0	227.7	227.7
SD:		1.00	4.73	14.47

 

Table 5: Summary of Body Weight Gains (g)

 

FEMALES		Day 0-7	Day 7-15	Day 0-15
Group 1: Solubilised Sulphur Black 1
2000 mg/kg bw, Step 1
Group size:		3	3	3
Mean: (g)		27.3	17.0	44.3
SD:		6.35	6.56	2.52
FEMALES		Day 0-7	Day 7-15	Day 0-15
Group 2:  Solubilised Sulphur Black 1
2000 mg/kg bw, Step 2
Group size:		3	3	3
Mean: (g)		26.7	0.0	26.7
SD:		4.62	19.16	14.57

Interpretation of results:

GHS criteria not met

Remarks:

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

Conclusions:

In an acute oral toxicity assay (acute toxicity class method, ATC) according to OECD guideline 423, an LD50 of above 2000 mg/kg bw was determined.

Executive summary:

The acute oral toxicity of the test item was determined in an in vivo toxicity test according to OECD guideline 423. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three young (8 weeks) female Wistar rats. No animal died in the first step at 2000 mg/kg bw, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too. Since the stopping criteria of Annex 2d of OECD Guideline No. 423 were met, the test was finished.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on the 15th day after the treatment.

No mortality was noted at a single oral dose of 2000 mg/kg bw. In the first step and second step, disturbance of the autonomic functions (diarrhoea) was observed in animals on the treatment day between 1 and 4 hours after the treatment. Black coloured faces/diarrhea observed was related to the physical property of the test item.

A body weight loss was observed in one female of group 2 in the second week. This body weight loss was not strong (ca. 9.7 %) and the animal gained body weight until the end of the study, thus it can be assessed as an individual variation without toxicological meaning. The mean body weight of both groups corresponded to their species and age throughout the study. All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

The method used is not intended to allow the calculation of a precise LD50value. Therefore, the test item was ranked into classes of the Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423. According to the classification system of the GHS, a LD50 of above 2000 mg/kg bw was determined.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Guideline and GLP conform study.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Oral

The acute oral toxicity of the test item was determined in an in vivo toxicity test according to OECD guideline 423. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three young (8 weeks) female Wistar rats. No animal died in the first step at 2000 mg/kg bw, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too. Since the stopping criteria of Annex 2d of OECD Guideline No. 423 were met, the test was finished.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on the 15th day after the treatment. No mortality was noted at a single oral dose of 2000 mg/kg bw. In the first step and second step, disturbance of the autonomic functions (diarrhoea) was observed in animals on the treatment day between 1 and 4 hours after the treatment. Black coloured faces/diarrhea observed was related to the physical property of the test item. A body weight loss was observed in one female of group 2 in the second week. This body weight loss was not strong (ca. 9.7 %) and the animal gained body weight until the end of the study, thus it can be assessed as an individual variation without toxicological meaning. The mean body weight of both groups corresponded to their species and age throughout the study. All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

The method used is not intended to allow the calculation of a precise LD50 value. Therefore, the test item was ranked into classes of the Globally Harmonized Classification System (GHS) described in the OECD guideline 423. According to the classification system of the GHS, a LD50 of above 2000 mg/kg bw was determined.

Dermal/Inhalation:

The in vivo study on acute dermal and inhalation toxicity was waived in accordance with Regulation (EG) No 1907/2006, Annex VIII, column 2.

The substance is highly hydrophilic (water solubility 617 g/L, log Pow < -3) and has a mean MW > 600000 Da with no fraction below 9500 Da. These substance characteristics severely restrict its systemic bioavailability (see also Toxicokinetic section). Considering the above and the result of the acute oral toxicity study, it can be concluded in a weight of evidence approach that the dermal LD50 is > 2000 mg/kg bw and the inhalation LC50 is > 5 mg/L.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available data are reliable and suitable for classification purposes under Regulation 1272/2008 (CLP). As a result the test item does not need to be classified for acute toxicity under Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EU) No 2019/521.