Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam distillation concentrate

Endpoint summary

Description of key information

The test item Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes steam distillation condensate (EC 942-492-8) was evaluated for acute oral toxicity in Sprague Dawley rats according to the OECD TG 420, GLP compliant.
A starting dose of 300 mg/kg body weight was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg body weight because of unavailability of sufficient toxicological data on the test item.
A total of 6 females were used for the experiment. All the animals of Sighting Study I, Sighting Study II and Main Study were administered with 300 mg/kg, 2000 mg/kg and 2000 mg/kg body weight respectively of the test item through oral route.

No clinical signs of toxicity and mortalities were observed in any of the dosed animals.
No changes were observed in body weight and percent change in body weight with respect to day 1. All the animals revealed physiologically normal increase in the body weight.
No gross pathological changes were observed in any of the dosed animals.

Based on the results of the experiment, it is concluded that the LD50 value for the test item is >2000 mg/kg body weight and, therefore, the substance is not classified as acutely toxic according to CLP criteria (Regulation
(EC) No 1272/2008).

 

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:

24 May - 19 June 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

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:

Batch No: 02 November 2020

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: In-house bred animals
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 161.15 g to 178.85 g
- Fasting period before study: fasted overnight (16 to 18 hours)
- Housing: Three animals were housed in standard polysulphonate cage (Size: L 430 x B 280 x H 210 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized paddy husk was provided as bedding material.

- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Healthy young adult animals used for Sighting Study I, Sighting Study II and Main Study were acclimatized for five, seven and nine days respectively to laboratory condition prior to treatment and were observed for clinical signs once daily.
- Method of randomisation in assigning animals to test and control groups

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6°C to 22.8°C
- Humidity (%): 46% to 64%
- Air changes (per hr): 12 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours fluorescent light and 12 hours dark cycle.

IN-LIFE DATES: From: 19 May To: 7/06/2021

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 and 200 mg/mL
- Amount of vehicle (if gavage): 5 ml
- Justification for choice of vehicle: As per the in-house miscibility test, test item is miscible in corn oil. Corn oil is universally accepted and routinely used vehicle in oral toxicity studies.
- Lot/batch no. (if required): N12009002


MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

- Rationale for the selection of the starting dose: A starting dose of 300 mg/kg body weight was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg body weight because of unavailability of sufficient toxicological data on the test item

Doses:

The test item was administered through oral gavage as a single dose of 300 mg/kg body weight to one female rat in Sighting study I. No clinical signs of toxicity and mortality was observed. Hence, as per the decision rules governing the sequential procedure presented in the OECD 420 test guideline, Sighting Study II was conducted using one female rat after approximately 24 hours of observation by administering a single dose of 2000 mg/kg body weight of the test item. No clinical signs of toxicity and mortality was observed. Hence, Main study was conducted using four female rats after approximately 24 hours of observation by administering a single dose of 2000 mg/kg body weight of the test item. No clinical signs of toxicity and mortality was observed.
Dose levels higher than 2000 mg/kg body weight were not tested.

No. of animals per sex per dose:

1 animal for sighting study at 300 mg/kg bw
5 animals for main studyat 2000 mg/kg bw
Total of 6 females were received.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All the animals were observed for clinical signs of toxicity and mortality at 20 to 30 mins, 1 hr (±10 mins), 2 hrs (±10 mins), 3 hrs (±10 mins) and 4 hrs (±10 mins) post dosing on day 1 and once daily thereafter for clinical signs of toxicity and twice daily for mortality during the 14 days observation period.
- Necropsy of survivors performed: yes, at the end of observation period, all the surviving animals were sacrificed under carbon dioxide asphyxiation, subjected to necropsy and a complete gross pathological examination and the observations were recorded. Observations included changes in skin, fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern.
- Clinical signs including body weight : Individual animal body weight was recorded at receipt, on day 1 (before test item administration) and on days 8 and day 15 during the observation period.
- Other examinations performed: Histopathological examination was not carried out as there were no gross lesions observed.

Preliminary study:

The test item was administered through oral gavage as a single dose of 300 mg/kg body weight to one female rat in Sighting study I. No clinical signs of toxicity and mortality was observed.
,Sighting Study II was conducted using one female rat after approximately 24 hours of observation by administering a single dose of 2000 mg/kg body weight of the test item. No clinical signs of toxicity and mortality was observed.
Main study was conducted using four female rats after approximately 24 hours of observation by administering a single dose of 2000 mg/kg body weight of the test item. No clinical signs of toxicity and mortality was observed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

other: No clinical signs of toxicity and mortality were observed in any of the dosed animals.

Gross pathology:

No gross pathological changes were observed in any of the dosed animals.

Other findings:

N/A

TABLE 1. INDIVIDUAL ANIMAL CLINICAL SIGNS OF TOXICITY AND MORTALITY RECORD

Study Steps & Dose (mg/kg body weight)

Animal No.

Sex

Time of Dosing (AM)

Clinical Signs of Toxicity and Mortality on Day 1

Clinical Signs of Toxicity and Mortality on Day

20 to 30 min

1 hr (±10 min)

2hrs (±10 min)

3hrs (±10 min)

4hrs (±10 min)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Sighting Study I                     &                          300

Rf6896

F

11:16

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Sighting Study II                   &                       2000

Rf6897

F

11:08

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Main Study      &                      2000

Rf6898

F

10:53

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rf6899

F

10:53

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rf6900

F

10:53

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rf6901

F

10:54

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N: Normal; F: Female; min/mins: minutes; hr/hrs: Hour/Hours

 

TABLE 2. INDIVIDUAL ANIMAL BODY WEIGHT (g) AND PERCENT CHANGE IN BODY WEIGHT WITH RESPECT TO DAY 1

Study Steps & Dose (mg/kg body weight)	Animal No.	Sex	Volume Administered (mL)	Body Weight (g) on Day			Percent Change in Body Weight with Respect to Day
				1	8	15		1 to 8	1 to 15
Sighting Study I &                                300	Rf6896	F	1.6	158.00	181.06	204.18		14.59	29.23
Sighting Study II &                             2000	Rf6897	F	1.6	155.78	175.10	193.02		12.40	23.91
Main Study                      &                            2000	Rf6898	F	1.6	157.02	170.15	192.09		8.36	22.33
	Rf6899	F	1.6	163.99	184.98	201.74		12.80	23.02
	Rf6900	F	1.6	160.18	178.86	205.15		11.66	28.07
	Rf6901	F	1.6	156.87	175.35	198.83		11.78	26.75
			Mean	159.52	177.34	199.45		11.15	25.04
			±SD	3.35	6.23	5.55		1.93	2.80

F: Female; SD: Standard Deviation

 

TABLE 3. INDIVIDUAL ANIMAL GROSS PATHOLOGY FINDINGS

Study Steps & Dose (mg/kg body weight)	Animal No.	Sex	Fate	Gross Pathology Findings
				External	Internal
Sighting Study I                              &                                 300	Rf6896	F	TS	NAD	NAD
Sighting Study II                             &                                       2000	Rf6897	F	TS	NAD	NAD
Main Study                      &                                      2000	Rf6898	F	TS	NAD	NAD
	Rf6899	F	TS	NAD	NAD
	Rf6900	F	TS	NAD	NAD
	Rf6901	F	TS	NAD	NAD

NAD: No Abnormality Detected; F: Female; TS: Terminal Sacrifice

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Based on the results of the experiment, it is concluded that the LD50 value for the test item Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes steam distillation condensate (EC 942-492-8) is >2000 mg/kg body weight when administered as a single dose by oral gavage to female Sprague Dawley rat as per OECD Guidelines for Testing of Chemicals No. 420 (Section 4: Health Effects) on conduct of “Acute Oral Toxicity - Fixed Dose Method”. The substance is unclassified as per CLP regulation.

Executive summary:

The test item Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes steam distillation condensate (EC 942-492-8) was evaluated for acute oral toxicity in Sprague Dawley rats according to the OECD TG 420. The study was GLP compliant.
A starting dose of 300 mg/kg body weight was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg body weight because of unavailability of sufficient toxicological data on the test item.
A total of 6 females were used for the experiment. All the animals of Sighting Study I, Sighting Study II and Main Study were administered with 300 mg/kg, 2000 mg/kg and 2000 mg/kg body weight respectively of the test item through oral route.

No clinical signs of toxicity and mortalities were observed in any of the dosed animals.
No changes were observed in body weight and percent change in body weight with respect to day 1. All the animals revealed physiologically normal increase in the body weight.
No gross pathological changes were observed in any of the dosed animals.

Based on the results of the experiment, it is concluded that the LD50 value for the test item is >2000 mg/kg body weight  via the oral route, therefore, the substance is not classified as acutely toxic according to CLP criteria (Regulation
(EC) No 1272/2008).

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

other: Expert Assessment

Adequacy of study:

key study

Study period:

2021

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: An assessment of the known constituents of the substance was performed to identify those associated with known acute toxicity and STO-SE properties via the inhalation route.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The acute toxicity potential and STOT-SE effects of Thermal cracking oil from blends of
rubber, fuel oils and paraffin waxes, steam distillation condensate was evaluated by assessing
the mixture in order to identify the presence of constituents known to be acutely toxic or have
specific target organ effects - single exposure (STOT-SE) via the inhalation route.

GLP compliance:

no

Test type:

other: Expert Assessment

Limit test:

no

Dose descriptor:

other: An assessment of the known constituents of the substance was performed to identify those associated with known acute toxicity and STO-SE properties via the inhalation route.

Remarks on result:

other: An assessment of the known constituents of the substance was performed to identify those associated with known acute toxicity and STO-SE properties via the inhalation route.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

It can be concluded that Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam distillation condensate shall be classified as Acute Toxic Category 4
with the Hazard Statement H332: Harmful if inhaled and as STOT-SE Category 3 with the Hazard Statement H336: May cause drowsiness or dizziness.

Executive summary:

The acute toxicity potential and STOT-SE effects of Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam distillation condensate was evaluated by assessing the mixture in order to identify the presence of constituents known to be acutely toxic or have specific target organ effects - single exposure (STOT-SE).

This assessment allowed us to identify 3 constituents (i.e. Ethylbenzene, Styrene and pXylene), classified as Acute Toxic Category 4 via the inhalation route, each present above the 1% cut-off level and 55.33% of other ingredients of unknown toxicity. As such, the equation for the Acute Toxicity Estimate (ATE) Method for classifying a mixture for its acute toxicity when a mixture contains more than 10% of ingredients with unknown toxicity was used.
The calculated ATE of the mixture gave a value of 24.28 mg/ l and was therefore found above the upper limit for Category 4 (20 mg/l). However, since the relevant constituents of the mixture are in a concentration range between ≥ 0.001m - ≤ 20%, we decided to use a worst-case scenario and classify the mixture for Acute Toxicity Category 4 - H332.

Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam distillation
condensate does not contain ingredients classified as STOT-SE Category 1 or 2, however, the mixture does contain above 20% sum of the ingredients classified as STOT-SE category 3 - H336. As such, classification of the mixture as STOT-SE Cat. 3 - H336 is applicable.

It can therefore be concluded that Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam distillation condensate shall be classified as Acute Toxic Category 4 with the Hazard Statement H332: Harmful if inhaled and as STOT-SE Category 3 with the Hazard Statement H336: May cause drowsiness or dizziness.

This assessment was performed using valid available data on all the components of the registered substance, and is considered as sufficient in order to conclude on its classification for acute toxicity via the inhalation route, using the rules laid down in the CLP Regulation. Further testing is not considered required

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Quality of whole database:

The acute toxicity potential and STOT-SE effects of Thermal cracking oil from blends of
rubber, fuel oils and paraffin waxes, steam distillation condensate was evaluated by assessing
the mixture in order to identify the presence of constituents known to be acutely toxic or have
specific target organ effects - single exposure (STOT-SE) via the inhalation route.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because inhalation of the substance is likely

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The LD50 value for the test item is >2000 mg/kg body weight and therefore, the substance is not classified as acutely toxic according to CLP criteria (Regulation
(EC) No 1272/2008).