L-arabinose

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-06-04 to 2018-06-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

17 July 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

May 30, 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No. of test material: AD16081001
- Expiration date of the batch: 2019-08-09

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under test conditions: Stable under ambient conditions

Analytical monitoring:

yes

Details on sampling:

- Sampling method: five replicate samples (2 mL per replicate) were taken from the test solution (100 mg/L) and one from the control (2 mL per replicate) at the start and at the end of the test.
- Sample storage conditions before analysis: Analyzed immediately after sampling

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The ISO medium was prepared by adding 25 mL from each of four stock solutions to one litre deionised water. The test solution used in the test was prepared by mechanical dispersion without using of any solubilising agent.
- Controls: Negative control, only ISO Test Medium without the test item
- Evidence of undissolved material: No

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: Akvárium Magazin Kft. (Pasaréti Gyula), 1222 Budapest, Dévény u. 36., Hungary
- Number of animals: 10 animals at each group (no replicates used)
- Age and size of animals: Juveniles; size within a range of 2.0 ± 1 cm
- Loading: The loading of the test aquaria did not exceed 1.0 g / L
- Sex: Both female and male (not separated)
- Animal health: Apparently healthy animals were used in the test

ACCLIMATION
Fish were held for at least 12 days before test initiation in the fish laboratory under the same conditions as used during the exposure period. During holding, fish were fed with appropriate, commercial diet for fish at least three times per week until 24 hours before the test started. The fish were not fed during the test. The health of the breeding was continuously monitored and any mortality or abnormal behaviour was recorded.
The mortalities were recorded for seven days immediately before the test and the following criteria were applied:
- Mortalities of greater than 10 per cent of population in seven days: Rejection of entire batch
- Mortalities of between 5 and 10 per cent of population: Acclimatisation continued for seven additional days
- Mortalities of less than 5 per cent of population: Acceptance of batch

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

249 mg/L (as CaCO3)

Test temperature:

22.6 – 23.5 °C

pH:

7.07 – 7.87

Dissolved oxygen:

62.8 – 96.3 % of the air saturation value at the temperature used

Salinity:

Not applicable

Conductivity:

Not specified

Nominal and measured concentrations:

Nominal concentration: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquariums with 5 litre test liquid
- Aeration: test solutions were not aerated during the test
- No. of organisms per vessel: 10
- No. of vessels per concentration: 1
- No. of vessels per control: 1

TEST MEDIUM
- Preparation of dilution water: ISO Medium (according to guideline)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light: 8 h dark

EFFECT PARAMETERS MEASURED: mortality, signs of intoxication

TEST CONCENTRATIONS
- Test concentrations in the main test: Based on the results of the non-GLP preliminary experiment the test item had no toxic effects on Zebrafish up to a concentration of 100 mg/L (limit concentration). Therefore, a limit test was performed using only the concentration of 100 mg/L. Concurrent control group ran.
- Pre-Experiment: A non-GLP preliminary experiment was carried out at 100 mg/L nominal concentration to check whether the test item had any toxic effects up to the limit concentration (i.e. 100 mg/L).
- Preparation of test solutions: The test solution used in the main test was prepared by mechanical dispersion without using of any solubilising agent. An amount of 0.5005 g test item was dissolved in 5000 mL dilution water (ISO Medium) to obtain the nominal concentration of 100 mg/L.
Untreated control ran parallel in the test. After the formulation procedure the test animals were immediately introduced into test solution.

Reference substance (positive control):

no

Remarks:

not applicable

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There were no mortalities or any sub-lethal effects observed in 10 fish in the control group or in 10 fish exposed to the 100 mg/L test concentration during the 96-h test period.

Results with reference substance (positive control):

Positive control not conducted.

Reported statistics and error estimates:

Since a limit test was performed (and mortality was not occurred), no statistical analysis was necessary. The LC0, LC50, NOEC, LOEC and the LC100 values were determined directly from the raw data.

Sublethal observations / clinical signs:

Table 1: Cumulative Mortality Data

Test Group	Cumulative mortality (initial population = 10 fish / test group)
	5h	24h	48h	72h	96h
Control	0	0	0	0	0
100 mg/L	0	0	0	0	0

Table 2: Measured and calculated Body Weight

Test Group	Measured weight of 10 fish (g)	Calculated mean weight of 1 fish (g)	Loading of testing aquarium (g fish/L testing liquid)
Control	2.01	0.201	0.40
100 mg/L	2.11	0.211	0.42

Table 3: Body Length of Test Animals

Test Group	Body length [cm]
Control	2.35	2.30	2.55	2.35	2.50	2.40	2.25	2.45	2.40	2.50
100 mg/L	2.45	2.40	2.45	2.40	2.45	2.45	2.30	2.35	2.35	2.45

Table 4: pH-values measured during the Experiment

Test Group	pH
	0h	24h	48h	72h	96h
Control	7.87	7.75	7.61	7.66	7.82
100 mg/L	7.82	7.65	7.07	7.38	7.48

Table 5: Dissolved Oxygen Concentrations measured during the Experiment

Test Group	Dissolved Oxygen Concentration[%]
	0h	24h	48h	72h	96h
Control	96.3	84.5	86.3	80.8	84.0
100 mg/L	95.8	78.8	67.8	65.1	62.8

Table 6: Temperatures measured during the Experiment

Test Group	Temperature[°C]
	0h	24h	48h	72h	96h
Control	22.6	23.3	23.4	23.2	23.4
100 mg/L	22.8	23.4	23.5	23.4	23.5

Table 7: Concentration of Test Item measured in the Test Solutions

Nominal concentration mg/L	Start (June 04, 2018)		End (June 08, 2018)
	Concentration (mg/L)	% of the nominal concentration	Concentration (mg/L)	% of the nominal concentration
Control	not detected	-	not detected	-
100	105	105	89.8	90
	106	106	88.7	89
	110	110	88.5	88
	107	107	88.3	88
	104	104	87	87
Mean:	106	106	88.5	88
RSD:	2	2	1	1

Validity criteria fulfilled:

yes

Conclusions:

In a GLP-compliant Acute Toxicity Test with Danio rerio according to OECD guideline 203, EU-Method C.1 and EPA OPPTS 850.1075, the 96-hour LC50 of the test item was determined to be > 100 mg/L (nominal).

Executive summary:

The acute toxicity of the test item to zebrafish (Danio rerio) was assessed in a 96-hour limit test under GLP conditions according to OECD guideline 203, EU-Method C.1 and EPA OPPTS 850.1075. Ten fish were exposed to aqueous test media containing the test item at the concentration of 100 mg/L. In addition, a negative control (ISO medium only) with 10 animals was performed. No replicates both for the limit concentration and the negative control were used. Fish were observed at approximately 5, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and temperature were carried out daily. Concentrations of the test item were determined at the start and the end of the test using the LC/MS/MS method.The measured concentrations of the test item were in the range of 104 – 110 % of the nominal concentration at the start of the test and were in the range of 87– 90 % of the nominal concentration at the end of the test. Therefore, biological results are based on nominal test item concentrations and endpoints. As a limit test was performed, effect values were determined directly from the raw data. In result, the test item had no toxic effect on Danio rerio at 100 mg/L concentration (i.e. limit test concentration). Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L. All validity criteria of the guidelines were met.

Description of key information

In an Acute Toxicity Test with Danio rerio according to OECD TG 203 the 96-hour LC50 of the substance was determined to be > 100 mg/L.

Additional information

The acute toxicity of the test item to zebrafish (Danio rerio) was assessed in a 96-hour limit test under GLP conditions according to OECD guideline 203, EU-Method C.1 and EPA OPPTS 850.1075. Ten fish were exposed to aqueous test media containing the test item at the concentration of 100 mg/L. In addition, a negative control (ISO medium only) with 10 animals was performed. No replicates both for the limit concentration and the negative control were used. Fish were observed at approximately 5, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and temperature were carried out daily. Concentrations of the test item were determined at the start and the end of the test using the LC/MS/MS method.The measured concentrations of the test item were in the range of 104 – 110 % of the nominal concentration at the start of the test and were in the range of 87– 90 % of the nominal concentration at the end of the test. Therefore, biological results are based on nominal test item concentrationsand endpoints. As a limit test was performed, effect values were determined directly from the raw data. In result, the test itemhad no toxic effect on Danio rerio at 100 mg/L concentration (i.e. limit test concentration).Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L. All validity criteria of the guidelines were met.