Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2021

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Reference substance name:
Low molecular weight organic substances from fast pyrolysis bio-oil
IUPAC Name:
Low molecular weight organic substances from fast pyrolysis bio-oil
Constituent 2
Reference substance name:
Monomers from fast pyrolysis bio-oil
IUPAC Name:
Monomers from fast pyrolysis bio-oil
Constituent 3
Reference substance name:
Dimers from fast pyrolysis bio-oil
IUPAC Name:
Dimers from fast pyrolysis bio-oil
Constituent 4
Reference substance name:
Trimers from fast pyrolysis bio-oil
IUPAC Name:
Trimers from fast pyrolysis bio-oil
Constituent 5
Reference substance name:
Higher oligomers from fast pyrolysis bio-oil
IUPAC Name:
Higher oligomers from fast pyrolysis bio-oil
Constituent 6
Chemical structure
Reference substance name:
Water
EC Number:
231-791-2
EC Name:
Water
Cas Number:
7732-18-5
Molecular formula:
H2O
IUPAC Name:
water
Test material form:
liquid: viscous
Remarks:
Brown /black highly vicous liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Institute of Experimental Pharmacology and Toxicology, Center of Experimental Medicine of the Slovak Academy of Sciences, Dobrá Voda, Slovak Republic
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 210g, 210g, 209g, 230g, 235g, 245g
- Fasting period before study: Animals were fasted prior to dosing food but not water was withheld over-night))
- Housing: The animals were housed in plastic cages suspended on stainless steel racks, up to 3 animals per cage, in a room equipped with central air- conditioning.
- Diet (e.g. ad libitum): A standard laboratory food KKZ-P/M (UEFT CEM SAS) was available ad libitum.
- Water (e.g. ad libitum): The animals received tap water for human consumption. Supply of drinking water was unlimited.
- Acclimation period: The animals were acclimated to the condition identical to the condition during the experiment at least 5 days prior to the start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): The room temperature was maintained within the range of 22±2 °C.
- Humidity (%): The relative humidity was 55±10 %.
- Photoperiod (hrs dark / hrs light): The light regime was set to a 12-hour light / 12-hour dark cycle.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 96% Ethyl alcohol p.a. diluted to 36% with Aqua pro Injectione
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2000 mg of the test item
- Amount of vehicle (if gavage): 36% ethanol (3 ml)
- Justification for choice of vehicle: Aqueous or oil solution/suspension/emulsion was not possible to prepare in order to administer proper dose to animals. BTG Pyrolytic Lignin was dissolved in in 96% Ethanol and further diluted with Aqua pro Injectione to the concentration of 36%. Further dilution caused precipitation of tested substance.
- Lot/batch no. (if required): Ethyl alcohol – 12.11.2019; Aqua pro Injectione – 19L1001

CLASS METHOD
- Rationale for the selection of the starting dose: Available information indicated that the test item was likely to be non-toxic regarding acute toxicity therefore we chose a dose of 2000 mg of BTG Pyrolytic Lignin per kg body weight to be used as a starting dose.
Doses:
2000 mg/kg body weight mixed with the vehicle 36% ethanol (3 ml).
No. of animals per sex per dose:
6 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: The animals were observed individually immediately after the administration of the test item and then 0.5, 1, 2 and 4 hours later. Each animal was inspected daily for the next 14 days.
- Frequency of observations and weighing: Individual weights of animals were determined shortly before the test item administration and weekly thereafter.
- Necropsy of survivors performed: All test animals were subjected to gross necropsy
- Other examinations performed: Examinations included external body surface and orifices, the appearance of tissues and organs in the thoracic cavity (trachea, esophagus, heart, aorta, lungs with main stem bronchi, thymus, tracheobronchial lymph node) and in the abdominal cavity (liver, spleen, adrenal glands, kidneys, ovaries, uterus including cervix, urinary bladder, small intestine, large intestine, pancreas, stomach, mesenteric lymph nodes).
Statistics:
not specified

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
other: Mortality
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed during the study.
Clinical signs:
other: No signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 14-days observation period
Gross pathology:
No visible pathological findings were observed.

Any other information on results incl. tables

Summary results of body weight

Sex

Dose

ID

Body Weight (g)

Body Weight Difference (g)

Initial

Week 1

Week 2

Week 1 - Initial

Week 2 - Initial

Week 2 – Week 1

Female

2000 mg/kg

1

210

231

237

21

27

6

2

210

216

228

6

18

12

3

209

243

254

34

45

11

4

230

246

258

16

28

12

5

235

250

262

15

27

12

6

245

256

271

11

26

15

Necropsy Results

Sex

Dose

ID

Result

Female

2000 mg/kg

1

no findings

2

no findings

3

no findings

4

no findings

5

no findings

6

no findings

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The LD50 of the test item is greater than 2000 mg/kg body weight after single oral administration to Wistar rats. No effects were observed at the limit dose of 2000 mg/kg bw.
Executive summary:

The purpose of the study was to evaluate the potential toxic effect of the test item when administered as a single oral dose to Wistar rats.
The procedure according to OECD Guideline 423 Acute Toxic Class (ATC) method was used. Available information indicated that the test item is likely to be non-toxic regarding acute toxicity, therefore, a limit dose of 2000 mg/kg body weight was used as a starting dose. The test item at this dose did not cause death in the next 48 hours and therefore another 3 females were treated at the same dose level. Pyrolytic lignin did not induce signs of intoxication, change of health, nor any other adverse reactions during 14-days observation period. During necropsy it was not observed any macroscopic findings in any of the animals.

It was concluded that the LD50 of the test item Pyrolytic lignin is greater than 2000 mg/kg body weight after single oral administration to Wistar rats.