Propionamide

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:

2020-08-26 to 2021-01-25

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:

18 June 2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 100 mg/L
- Sampling method: duplicate samples from the freshly prepared test media of the only concentration and the control were taken at the start of the test and at day 3. For the determination of the stability of the test item under the test conditions, respectively the maintenance of the test item concentration during the test period, samples were taken in duplicate out of the test medium and the control at day 1 and at the end of the test at day 4 from the approximate centre of the aquaria.
The pH value was adjusted to pH 3 using hydrogen chloride directly after sampling.
- Sample storage conditions before analysis: all samples were stored in a freezer (≤ -20 °C), protected from light until analysis was performed

Vehicle:

yes

Remarks:

Reconstituted Water

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The test medium of the only test item concentration of nominal 100 mg/L was prepared by dissolving 1091.0, 1082.5, 1096.8 and 1106.4 mg test item into 1091.0, 1082.5, 1096.8 and 1106.4 mL test water by intense stirring for 5 minutes. The test media was prepared just before introduction of the test fish (= start of the test and test medium renewal on day 1, 2 and 3).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: the test fish were obtained from in house breeding
- Length at study initiation: the mean body length of the fish in the test was 1.57 cm ± 0.1 cm (Mean ± SD),
- Weight at study initiation: the mean body wet weight was 0.065 g ± 0.02 g (Mean ± SD),
- Feeding: three times per week or daily

ACCLIMATION
- Acclimation period: All fish were obtained and held in the laboratory for at least 9 days before the start of the test. They were held in water of the quality to be used in the test for at least seven days immediately before testing under the following conditions:
Light: 12 hours photoperiod daily
Temperature: 21- 25 °C
Oxygen concentration: at least 80 % of the air saturation value
- Acclimation conditions: same as test
- Feeding frequency during acclimation: three times per week or daily until 24 hours before the test was started
- Health during acclimation (any mortality observed): No

FEEDING DURING TEST: No

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

1.78 mmol/L (= 178.4 mg/L) as CaCO3

Test temperature:

22.3 - 22.9 °C

pH:

7.3 - 7.9

Dissolved oxygen:

99 - 103 % of the air saturation value

Details on test conditions:

TEST SYSTEM
- Test vessel: 12 L glass aquaria with 10 L test medium
- Type: open
- Aeration: the test media were slightly aerated during the test
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water according to OECD 203
- Ca/mg ratio: 4:1
- Culture medium different from test medium: no
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12 hour light and 12 hour dark regime

EFFECT PARAMETERS MEASURED: mortality and sublethal effects

TEST CONCENTRATIONS
- Limit test
- Test concentrations: 100 mg/L and 0 mg/L (control)

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)

Details on results:

For details please refer to "Any other information on results". No further effects were observed.

Sublethal observations / clinical signs:

Biological Results

Mortality and sublethal effects: In the control and the only test concentration of 100 mg test item/L , all fish survived until the end of the experiment and showed no sublethal effects during the exposure time.

 

Table 1. Observed Mortality of unfed Zebrafish (Danio rerio) exposed the test item for 96 hours

Nominal concentration	Mortality
[mg/L]	0 h	24 h	48 h	72 h	96 h
	# mort	# mort	# mort	# mort	# mort
Control	0	0	0	0	0
100	0	0	0	0	0
LC50 [mg/L]	-	> 100	> 100	> 100	> 100
95 % CI	-	n.d.	n.d.	n.d.	n.d.

 

96h LC50	> 100 mg test item/L
96h LC20	> 100 mg test item/L
96h LC10	> 100 mg test item/L
96h LC0	100 mg test item/L
96h LC100	> 100 mg test item/L
96h NOEC	≥ 100 mg test item/L
96h LOEC	> 100 mg test item/L

 

Analytical Results

Table 2. Summary of analytical results

Sample Description	fresh (0h)			aged (24h)
[mg test item/L]	% of nominal^1	RSD [%]	n	% of nominal^1	RSD [%]	n
Control	n.a.	n.a.	4	n.a.	n.a.	4
100	97	0.4	4	96	1	4

^1 mean value of all measured samples per treatment group RSD: relative standard deviation per treatment group n: number of analysed samples n.a.: not applicable

 

Validity of the Test

No mortality was seen during the acclimation phase. Therefore the mortalities in the fish batch were below 5 % and the fish batch was accepted.

The mortality in the control did not exceed 10 % at the end of the study.

The dissolved oxygen concentration was > 60 % throughout the study.

Thus, the validity criteria were fulfilled.

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to zebrafish Danio rerio was determined according to OECD TG 203. Based on the test results the 96-hour LC50 was determined to be > 100 mg test item/L based on the nominal concentration.

Executive summary:

The acute toxicity of the test item to zebrafish Danio rerio was determined in an aerated, semi-static limit test according to OECD TG 203. For this purpose, 7 zebrafish per test group and control were exposed over a 96 h period under defined conditions. The study comprised of one control group and one test item group. A nominal test item concentration of 100 mg/L was prepared in reconstituted water. The zebrafish were observed for signs of toxicity and mortality for 96 hours. The analytical determination of the test item concentration was performed using HPLC with UV-detection. Duplicate samples from the freshly prepared test media of the only concentration and the control were taken at the start of the test and at day 3. The limit of quantification (LOQ) of the analytical method was 50 mg/L. In the freshly prepared test media at the start of the test and at the renewal of the test media 97 % of the only test concentration were found. In the aged test media after 24 hours test duration, 96 % of the nominal value was determined. During the test the test organism were exposed to a mean of 96 % of nominal. In the control and the test concentration of 100 mg test item/L, all fish survived until the end of the experiment and showed no sublethal effects during the exposure time. The dissolved oxygen concentration in the test media did not fall below 99 % of air saturation value. As, no acute toxicity was observed at a nominal concentration of 100 mg/L, the 96 h LC50 was > 100 mg/L (nominal). The NOEC was determined to be > 100 mg test item/L also based on the nominal concentration.

Description of key information

The acute toxicity of the test item to zebrafish Danio rerio was determined according to OECD TG 203. Based on the test results the 96-hour LC50 was determined to be > 100 mg test item/L based on the nominal concentration (reference 6.1.1-1).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 100 mg/L

Additional information

The acute toxicity of the test item to zebrafish Danio rerio was determined in an aerated, semi-static limit test according to OECD TG 203. For this purpose, 7 zebrafish per test group and control were exposed over a 96 h period under defined conditions. The study comprised of one control group and one test item group. A nominal test item concentration of 100 mg/L was prepared in reconstituted water. The zebrafish were observed for signs of toxicity and mortality for 96 hours. The analytical determination of the test item concentration was performed using HPLC with UV-detection. Duplicate samples from the freshly prepared test media of the only concentration and the control were taken at the start of the test and at day 3. The limit of quantification (LOQ) of the analytical method was 50 mg/L. In the freshly prepared test media at the start of the test and at the renewal of the test media 97 % of the only test concentration were found. In the aged test media after 24 hours test duration, 96 % of the nominal value was determined. During the test the test organism were exposed to a mean of 96 % of nominal. In the control and the test concentration of 100 mg test item/L, all fish survived until the end of the experiment and showed no sublethal effects during the exposure time. The dissolved oxygen concentration in the test media did not fall below 99 % of air saturation value. As, no acute toxicity was observed at a nominal concentration of 100 mg/L, the 96 h LC50 was > 100 mg/L (nominal). The NOEC was determined to be > 100 mg test item/L also based on the nominal concentration.