Sodium hydrogen L-aspartate

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:

2017-08-28 to 2017-09-01

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:

adopted July 17, 1992

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

April 1986

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling method: five replicate samples (5 mL per replicate) were taken from the test solution (100 mg/L) and from the control at the start and at the end of each water renewal periods.
- Sample storage conditions before analysis: Analyzed immediately after sampling

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Separate stock solutions of individual substances were first prepared in deionised water. 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 (e.g. precipitate, surface film, etc.): 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
- Age at study initiation: Juveniles
- Length at study initiation: 2 ± 1 cm
- Maintenance of the brood fish: Appropriate, commercial diet for fish at least three times per week until one day before the test start

HOLDING AND ACCLIMATION
- Acclimation period: At least 12 days before test initiation
- Acclimation condition: Same as test
- Health: No significant mortality (less than 5% of the population) occurred in seven days before the start of the experiment



FEEDING DURING TEST: No

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No

Hardness:

249 mg/L CaCO3

Test temperature:

22.0 - 23.0 °C

pH:

7.44 – 7.80

Dissolved oxygen:

71.7 – 92.6 % of the air saturation value at the temperature used

Nominal and measured concentrations:

Nominal Concentration: 100 mg/L
Measured concentration: 97.95 mg/L at the start and 93.88 mg/L at the end of the test

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquariums with 3 litre test liquid
- Aeration: test solutions were not aerated during the test
- Renewal rate of test solution: at day 2 of the test
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.92 g/L in the control and 0.83 g/L in the test item treated group

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ultrapure water (ASTM Type I), prepared by Direct-Q 5 system, MILLIPORE

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

EFFECT PARAMETERS MEASURED: mortality, signs of intoxication

TEST CONCENTRATIONS
- Limit concentration: 100 mg/L
- Range finding study: yes, no toxic effects on Zebrafish up to a concentration of 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

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)

Remarks on result:

other: measured LC50 for hydrat

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: No
- Observations on body length and weight: No
- Other biological observations: No
- Mortality of control: No
- Other adverse effects control: No
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

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	Cummulative mortality (initial population = 10 fish / test group)
	4h	24h	48h	72h	96h
Control	0	0	0	0	0
100 mg/L	0	0	0	0	0

Table 2: Measured and calculated Data of Body Weight

Test group	Measured weight of 10 fish (g)	Calculated mean weight of fish (g)	Loading of testing aquarium (g fish/L testing liquid)
Control	2.76	0.28	0.92
100 mg/L	2.49	0.25	0.83

Table 3: Body Length of Test Animals

Test group	Body length [cm]
Control	2.30	2.50	2.30	2.30	2.20	2.25	2.50	2.25	2.45	2.40
100 mg/L	2.25	2.20	2.50	2.35	2.20	2.40	2.25	2.30	2.15	2.45

Validity criteria fulfilled:

yes

Conclusions:

The 96h-LC50 of the the test item was determined to be > 100 mg/L (nominal, hydrated substance). The test item had no toxic effect on fish at the limit concentration of 100 mg/L (nominal, hydrated substance). The 96-h LC50 and 96-h NOEC was calculated to be > 89.6 mg/L and 89.6 mg/L, respectively, for the anhydrous substance.

Executive summary:

The acute toxicity of the test substance to zebrafish (Danio rerio) was determined in a 96 -hour semi-static limit-test under GLP conditions according to OECD guideline 203, EU-Method C.1 and EPA OPPTS 850.1075. One single concentration of 100 mg/L was selected on the basis of a preliminary test. The test solution was prepared by mechanical dispersion without using of any solubilising agent and was renewed one during the test (at day 2). A negative control only with test medium (OECD Medium) ran in parallel. The vessel for the test solution and the control were loaded with 10 test organisms each. The measured concentrations of L-aspartic acid sodium salt monohydrate were in the range of 94 – 100 % of the nominal during the experiment. The measured test item concentration remained within ± 20 % of the nominal during the test period of 48 hours, therefore the biological results are based on the nominal concentration. The fish were observed at approximately 4, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. In result, the test item had no toxic effect on fish at 100 mg/L concentration (i.e. limit test concentration). Accordingly, the 96-h LC50 value was determined to be > 100 mg/L (nominal, hydrated substance). The 96-h NOEC was determined to be 100 mg/L (nominal, hydrated substance). The 96-h LC50 and 96-h NOEC was calculated to be > 89.6 mg/L and 89.6 mg/L, respectively, for the anhydrous substance. All validity criteria were met.

Description of key information

In a acute toxicity test with fish, conducted according to OECD TG 203, EU Method C.1 and OPPTS 850.1075, the 96h-LC50 of the test item was determined to be > 100 mg/L (reference 6.1.1 -1).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The acute toxicity of the test substance to zebrafish (Danio rerio) was determined in a 96 -hour semi-static limit-test under GLP conditions according to OECD guideline 203, EU-Method C.1 and EPA OPPTS 850.1075 (reference 6.1.1-1). One single concentration of 100 mg/L was selected on the basis of a preliminary test. A negative control only with test medium (OECD Medium) ran in parallel. The vessel for the test solution and the control were loaded with 10 test organisms each. The measured concentrations of the test item were in the range of 94 – 100 % of the nominal during the experiment. The fish were observed at approximately 4, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. In result, the test item had no toxic effect on fish at 100 mg/L (hydrat) concentration. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L (nominal, hydrated) and calculated as 89.6 mg/L

for the anhydrous substance. The 96-h NOEC was determined to be 100 mg/L (nominal, hydrated substance) and 89.6 mg/L (calculated for the anhydrous substance), respectively. All validity criteria were met.