5-(hydroxymethyl)-2-furaldehyde

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 May 2019 - 13 June 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

17th December 2001

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

67-47-0

Molecular formula:

C6H6O3

impurity 1

Reference substance name:

Fructose

EC Number:

200-333-3

EC Name:

Fructose

Cas Number:

57-48-7

Molecular formula:

C6H12O6

IUPAC Name:

D-fructose

impurity 2

Reference substance name:

1-(furan-2-yl)-2-hydroxyethanone

Cas Number:

17678-19-2

Molecular formula:

C6H6O3

IUPAC Name:

1-(furan-2-yl)-2-hydroxyethanone

impurity 3

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

impurity 4

Reference substance name:

Sodium formate

EC Number:

205-488-0

EC Name:

Sodium formate

Cas Number:

141-53-7

Molecular formula:

CH2O2.Na

IUPAC Name:

sodium formate

impurity 5

Reference substance name:

Sodium nitrate

EC Number:

231-554-3

EC Name:

Sodium nitrate

Cas Number:

7631-99-4

Molecular formula:

HNO3.Na

IUPAC Name:

sodium nitrate

impurity 6

Reference substance name:

Glucose

EC Number:

200-075-1

EC Name:

Glucose

Cas Number:

50-99-7

Molecular formula:

C6H12O6

IUPAC Name:

D-glucose

impurity 7

Reference substance name:

Sodium acetate

EC Number:

204-823-8

EC Name:

Sodium acetate

Cas Number:

127-09-3

Molecular formula:

C2H4O2.Na

IUPAC Name:

sodium acetate

impurity 8

Reference substance name:

2-furaldehyde

EC Number:

202-627-7

EC Name:

2-furaldehyde

Cas Number:

98-01-1

Molecular formula:

C5H4O2

IUPAC Name:

2-furaldehyde

impurity 9

Reference substance name:

Sodium 4-oxovalerate

EC Number:

243-378-4

EC Name:

Sodium 4-oxovalerate

Cas Number:

19856-23-6

Molecular formula:

C5H8O3.Na

IUPAC Name:

sodium 4-oxopentanoate

impurity 10

Reference substance name:

Formaldehyde

EC Number:

200-001-8

EC Name:

Formaldehyde

Cas Number:

50-00-0

Molecular formula:

CH2O

IUPAC Name:

formaldehyde

Test material form:

solid: crystalline

Details on test material:

freeze-dried material
Batch: 1808A-RO-C-I1-FD-Chg#3
Appearance Crystal / brown
Storage: Refrigerator (2 - 8 °C)
Expiring date: 31 December 2019

Specific details on test material used for the study:

Batch: 1808A-RO-C-I1-FD-Chg#3
Storage: Refrigerator (2 - 8 °C)

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST 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, recognized by international guidelines as the recommended test system (e.g. OECD, EC)
Number of animals: 3 animals/group; 6 in total
Sex:Female, nulliparous and non pregnant animals
Age of animals:Young adult rat, 9-11 weeks old in first and second step
Body weight range at starting (first step):181 - 183 g
Body weight range at starting (second step):185 - 195 g
Acclimatization time:19 days in first step and 20 days in second step.

ENVIRONMENTAL CONDITIONS
Animal health: Only healthy animals were used for the study. The health status of the animals used in this study was guaranteed by the supplier.
Room: 13/1
Housing: Group caging (3 animals/cage)
Cage type:Type III polypropylene/polycarbonate.
Bedding: 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.

FOOD AND WATER SUPPLY
Animals received ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany and tap water from municipal supply, as for human consumptionfrom bottle ad libitum. The diet and drinking water are periodically analysed and are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
Name:aqua purificata
Batch number: 190101
Date of expiration:07.07.2019
Produced by: Magilab Kft.

Formulation: All doses were formulated in the vehicle (distilled water). The vehicle was selected based on trial preparations performed at the testing facility and on test item data supplied by the sponsor. The concentration of the formulations was adjusted to maintain a treatment volume of 10 mL/kg bw. The test item was applied in a concentration of 200 mg/mL. The correction factor was taken into consideration in the course of the making of solution.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

Justification of the doses
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 female rats. The absence or presence of mortality of animals doses at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration an severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423 was met.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

3 females per dose (first step)
3 females per dose (second step)

Control animals:

no

Details on study design:

Procedure: A single oral administration - followed by a fourteen-day observation period - was performed by gavage, using plastic feeding tubes. The day before treatment the animals were fasted. The food but not water was withheld overnight. Animals were weighed before the application and the appropriate volume will be applied accordingly. T he food was given back 3 hours after the treatment. The first group was observed for up to min. 24 hours before the next step was treated.
Mortality: 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.

General state, external appearance, behavior and clinical symptoms: 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 once each day for 14 days thereafter. 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.

Measurement of Body Weight: The body weights were recorded on day 0 (just before the treatment), on day 7 and on day 15 with a precision of 1 g.

Necropsy: At the end of the observation period all rats were sacrificed under isofluran anaesthesia and subjected to necropsy. 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.

Statistics:

No statistical analysis was performed.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No death occurred at 2000 mg/kg bw single oral dose of 5-(Hydroxymethyl)furfural. 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/kgbw dose clinical sign of reaction comprised of decreased activity (6 cases of 57 observations), closed eyes (2/57) and piloerection (2/57). Decreased activity (score -1) was observed in all animals. Closed

Gross pathology:

All animals survived until the scheduled necropsy on Day 15. Moderate hydrometra was observed in female No.: 4780 of group 1 in animal No.: 4789 of group 2, as well severe hydrometra was detected in animal No.: 4791 of group 2. Hydrometra is physiological finding and connected to the oestrus cycle of the animal. Nopathological changes were found related to the effect of the test item during the macroscopic examination of animals treated with 2000 mg/kg bw dose.

Other findings:

No death occurred after the single 2000 mg/kg bw oral dose of 5-(Hydroxymethyl)furfural. The clinical signs observed in both groups were related to the effect of the test item. There were no any related to the effect of the test item found in body weights and body weight gains during the study. Autopsy revealed no treatment related pathological changes.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The method used, was not intended for the precise calculation of a precise LD50 value. The oral LD50 value of the test item in Wistar rats was established to exceed 2000 mg/kg body weight.The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as below:
Dose (mg/kg bw): 2000
Mortality(dead/treated): 0/6
LD50 (mg/kg bw): = 5 000
GHScategory: 5 or unclassified

In conclusion, the LD50 of the test item 5-(Hydroxymethyl)furfural is higher than 5000 mg/kg bodyweight by oral route in the rat. Based on these results, the test item 5-(Hydroxymethyl)furfural does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of classification and Labelling of Chemicals (GHS) of the United Nations (2017) (including all amendment) and regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Executive summary:

General information:

The goal of this study was to assess the acute oral toxicity with 5-(Hydroxymethyl)furfural in the Wistar rat (following the Acute Toxic Class Method).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  female  rats,  administered  by  oral  gavage.  No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was  finished;  the  stopping  criteria  of  Annex  2d  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  on  the  15th  day  after  the  treatment in animals.

Lethality, Clinical symptoms and Body weight:

No  death  occurred  at  2000  mg/kg  bw single  oral  dose  of  5-(Hydroxymethyl)furfural.  All  female rats in step 1 and step 2 survived until the end of the 14-day observation period. In a first  step,  CNS  - and  emotion  symptoms    (decreased  activity,  closed  eyes)  anddisturbance of autonomic functions (piloerection) were observed in animals on the treatment day between 30 minutes and 1 hour after the treatment.  In a  second  step,  CNS  -  and  emotion  symptoms  (decreased  activity,  closed  eyes,  bedding  chewing),  disturbances  of  coordination  (abnormal  gait,  incoordination),  decreased  righting  reflex  and  disturbances  of  autonomic  functions  (diuresis,  piloerection)  were  observed  in  animals on the treatment day between 1 and 4 hours after the treatment. The body weight development was undisturbed in all animals.

Gross pathology:

All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

Evaluation:

The method used is not intended to allow the calculation of a precise LD50 value. The oral LD50  value  of  the  test  item  in  Wistar  rats  was  established  to  exceed  2000  mg/kg  body  weight.  The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as below:

Dose (mg/kg bw): 2000

Mortality(dead/treated):  0/6

LD50 (mg/kg bw):  = 5 000

GHScategory: 5 or unclassified

In  conclusion,  the  LD50  of  the  test  item  5-  (Hydroxymethyl)furfural  is   higher  than  5000 mg/kg bodyweight by oral route in the rat.Based  on  these  results,  the  test  item  5-(Hydroxymethyl)furfural  does  not  have  to  be  classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of classification and Labelling of Chemicals (GHS) of the United  Nations  (2017)  (including  all  amendment)  and  regulation  (EC)  No  1272/2008  on  classification,   labelling   and   packaging   of   substances   and   mixtures   (including   all   amendments).