5-(hydroxymethyl)-2-furaldehyde

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 June 2019 - 22 July 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 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

30 May 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The following test vessels were set up:
- Test solution (Tn); containing Daphnia medium with test item (four replicates with five Daphnia each)
- Blank control (Bn); containing pure Daphnia medium (four replicates with five Daphnia each)
- Since the test item is soluble, the test solutions were prepared by respective dilutions of a 100 mg/l stock solution (test item dissolved in test water) with test water, which were used as the test solutions in the toxicity test. Pure Daphnia medium served as blank controls.
- O2 concentration and pH were measured in the test medium for each concentration including the blank control. If the O2 concentration was too low (>5.0 mg/l required) the media was aerated by stirring. Afterwards, the media were filled into the test vessels (50 ml per vessel).
- Daphnia, aged less than 24 h and already acclimatized to the Daphnia medium, were then introduced into the test media, and the vessels were covered with a glass plate. The Daphnia were not fed during the test and the test vessels were not aerated.
semi static conditions: After 24 h, the Daphnia were transferred to the fresh test media.
- Evidence of undissolved material: Not relevant. Since the test item is soluble, the test solutions were prepared by directly dissolving the test item in Daphnia medium

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISMS
- Test organism: Daphnia magna (Straus, 1820), derived from a healthy stock and not first brood progeny
- Breeding: Parental and young Daphnia held in 4 l glass aquaria (3 l medium) at 20±2°C
- Illumination: 16 h per day
- Medium: Continuously aerated Elendt M4 medium prepared with ultra-pure water (conductivity <1.5 µS/cm)
- Feed: Suspension of Desmodesmus subspicatus in Elendt M4 medium with an optical density OD680 of about 15 units

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

not reported

Test temperature:

18–22°C, controlled at ±1 °C

pH:

7.3 - 7.8

Dissolved oxygen:

The minimum dissolved oxygen concentration in the controls and the test vessels at the end of each 24 h-period was >=7.5 mg O2/l (required >=3 mg O2/l).

Salinity:

not reported

Conductivity:

670 µS cm-1

Nominal and measured concentrations:

The concentrations of 5-(Hydroxymethyl)furfural in the test medium were determined by HPLC analysis (LOQ 0.0129 mg/l) at the beginning, after 24 h in the old and the fresh media, and after 48 h of exposure. These analyses showed that the concentrations were correctly dosed and were within 80–120% of
the nominal value. Over the two test medium renewal periods of 24 hours however, the test item concentration in the test media decreased. At the end of the test medium renewal periods at 24h and 48 h, 37–59% and 55–70% of the initially measured concentration was found, respectively. Therefore, the effective concentrations ECx were assessed based on the geometric mean measured concentrations which were 74.2, 35.0, 16.2, 7.08, 2.98 and 1.19 mg/l for the 0–24 h interval, and 77.7, 35.2, 17.1, 7.50, 3.19 and 1.32 mg/l for the 0–48 h interval.

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml beakers, all-glass, with 50 ml of test medium,covered with a glass plate to avoid evaporation and contamination of the test solutions with dust
- Test medium: Elendt M4 medium; prepared with ultra-pure water (conductivity <1.5 µS/cm)
- Number of Daphnia: 20 individuals per test concentration, and for the blank, divided in 4 groups of 5 individuals
- Age: Less than 24 h
- Light: non-GLP preliminary test: 16 hours photoperiod per day, 550–1100 lux, supplied by overhead white fluorescent tubes
definitive test: in the dark
- Temperature: 18–22 °C, controlled at ±1 °C
- pH: 6 to 9. The pH should normally not vary by more than 1.5units in one test.
- Feed: The Daphnia are not fed during the test
- Aeration: The test vessels were not aerated during the test.
- Test type: Semi-static exposure conditions; renewal of test medium after 24 h of exposure
- Test duration: 48 h

RANGE-FINDING STUDY
Prior to the definitive test a non-GLP range finding test with nominal concentrations of 100, 10 and 1 mg/l of the test item was performed under static conditions (two replicates for each of the test concentration and the control; five daphnids per replicate) and normal light conditions (16 hours photoperiod per day, about 550–1100 lux, supplied by overhead white fluorescent tubes).

EFFECT PARAMETERS MEASURED
Observed immobility (inability to swim) of the daphnids: Observations of immobile Daphnia were made after 24 and 48 h of exposure. Any abnormal behaviour or appearance was reported.

Reference substance (positive control):

yes

Remarks:

Acute reference test with potassium dichromate conducted twice a year

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

46.5 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: calculated by linear regression

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

17.1 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: calculated by linear regression

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

71.7 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: calculated by linear regression

Effect concentrations 4

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

35 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: calculated by linear regression

Details on results:

After 24 hours of exposure the following immobilisation rates were observed: 60% at 74.2 mg/l. No significant effects (=10% immobilisation) were observed at 35.0, 16.2, 7.08, 2.98 and 1.19 mg/l or in the control.
The 24 h-EC50 of 5-(Hydroxymethyl)furfural to Daphnia magna was calculated to be 71.7 mg/l (95% confidence limits: 57.2–90.0 mg/l). This ECx value was computed using a linear regression with a logit link function and a maximum likelihood algorithm (slope = 27.6, Intercept = –51.3, degrees of freedom = 4, p(Chi²) <0.001, Chi2 =18.8, p(F) <0.001).
The no observed effect concentration (NOEC) after 24 hours was determined to be 35.0 mg/l.

After 48 hours of exposure the following immobilisation rates were observed: 100% at 77.7 mg/l and 15% at 35.2 mg/l. No significant effects (=10% immobilisation) were observed at 17.1, 7.50, 3.19 and 1.32 mg/l or in the control.
The 48 h-EC50 of 5-(Hydroxymethyl)furfural to Daphnia magna was calculated to be 46.5 mg/l (95% confidence limits: 36.1–59.8 mg/l). This ECx value was computed using a linear regression with a Weibull link function and a maximum likelihood algorithm (slope = 12.4, Intercept = –21.0, degrees of freedom = 4, p(Chi²) = 1.00, Chi2 =0.0630, p(F) <0.001).
The no observed effect concentration (NOEC) after 48 hours was determined to be 17.1 mg/l.

Results with reference substance (positive control):

Acute ref. test with K2Cr2O7 conducted twice a year. The EC50 value for the control of sensitivity for 24 h of exposure with K2Cr2O7 was estimated to be 0.89 mg/l (29.6.2016), which lies within the recomm. range of 0.6–2.1 mg/l acc. to OECD Guideline 202.

Reported statistics and error estimates:

The effective concentrations ECx were assessed based on the geometric mean measured concentrations which were 74.2, 35.0, 16.2, 7.08, 2.98 and 1.19 mg/l for the 0–24 h interval, and 77.7, 35.2, 17.1, 7.50, 3.19 and 1.32 mg/l for the 0–48 h interval.
The statistical analysis was conducted with the software ToxRat® Standard Version 3.2.1.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

All validity criteria i.e. immobilization in the controls <=10% and O2 concentrations at the end of the test >=3 mg/L were fulfilled.

Conclusions:

The acute toxicity of 5-(Hydroxymethyl)furfural (CAS no. 67-47-0) to Daphnia magna was determined in a 48 hour semi-static test according to OECD guideline 202. The median effect concentrations (EC50) of 5-(Hydroxymethyl)furfural on Daphnia magna after 24 h was calculated to be 71.7 mg/l (95% confidence limits: 57.2–90.0 mg/l) and after 48 h 46.5 mg/l (95% confidence limits: 36.1–59.8 mg/l). The NOEC values after 24 h was determined to be 35.0 mg/l and after 48 h of exposure 17.1 mg/l. The results of the test can be considered reliable without restriction.

Executive summary:

The acute toxicity of 5-(Hydroxymethyl)furfural (CAS no. 67-47-0) to Daphnia magna was investigated according to the OECD guideline 202 under semi-static exposure conditions over a period of 48 hours.

The test item 5-(Hydroxymethyl)furfural is solid, 85.4% pure and soluble in test water. Consequently, the test solutions were prepared by appropriate dilutions of a stock solution.

The nominal test item concentrations tested 100, 45.5, 20.7, 9.39, 4.27 and 1.94 mg/l. Four replicates for each test concentration as well as for the control, were tested.

The concentrations of 5-(Hydroxymethyl)furfural in the test media were analyzed by HPLC with UV detection at the beginning as well as after 24 hours in the fresh and old media and after 48 hours of exposure. These analyses confirmed that the concentrations were correctly dosed but that they decreased below 80% of the nominal value during the two test medium renewal periods of 24 hours. Therefore, the effective concentrations ECx were assessed based on the geometric mean measured concentrations which were 74.2, 35.0, 16.2, 7.08, 2.98 and 1.19 mg/l for the 0–24 h interval, and 77.7, 35.2, 17.1, 7.50, 3.19 and 1.32 mg/l for the 0–48 h interval.

After 24 hours of exposure, the following immobilisation rates were observed: 60% at 74.2 mg/l. No significant effects were observed at 35.0, 16.2, 7.08, 2.98 and 1.19 mg/l.

After 48 hours of exposure, the following immobilisation rates were observed: 100% at 77.7 mg/l and 15% at 35.2 mg/l. No significant effects were observed at 17.1, 7.50, 3.19 and 1.32 mg/l.

The biological test results of 5-(Hydroxymethyl)furfural to Daphnia magna after 24 and 48 hours of exposure are summarized in the following table based on the mean measured concentrations:

Parameter	Immobilisation (0-24 h) [mg/l]	Immobilisation (0-48 h) [mg/l]
EC50	71.7	46.5
NOEC	35.0	17.1

In conclusion, the 5-(Hydroxymethyl)furfural (CAS no. 67-47-0) had toxic effects on Daphnia magna. The 48-hour EC50 was estimated to be 46.5 mg/l.

All validity criteria were fulfilled.