Reaction mass of (2E)-Tridec-2-enenitrile and (2Z)-Tridec-2-enenitrile and (3E)-Tridec-3-enenitrile and (3Z)-Tridec-3-enenitrile

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2016-03-22 to 2017-01-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and all test item concentrations from the main test.
- Sampling method: Samples of the old media were taken directly (24 and 48 hours) from the test vessels.
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until the start of the analysis, if necessary. Prepared samples were stored in the autosampler at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A saturated solution of the test item was prepared two days prior to the start of the exposure intervals (at ~48 and ~24 hours) in a tightly closed glass flask. The glass flask was filled up with 2300 mL of the dilution water (ISO test water) and closed with a screw cap with a septum. For siphoning of the saturated solution, a glass tube was placed in the glass flask near the bottom and the septum was pierced. The test item (1 mL) was placed by pipette onto the water surface and the bottle was closed with the screw cap. A slow stirring procedure in a closed system without headspace was applied. Gentle stirring (to avoid formation of an emulsion) was carried out for 2 days with a magnetic stirrer at room temperature. After completion of the stirring period, the saturated solution was collected by siphon through the glass tube from the approximate bottom of the glass flask. The saturated solution was checked via laser beam (Tyndall effect) for undissolved test item (formation of an emulsion). Since a Tyndall effect was observed, the solution was filtered with a membrane filter (0.45 pm). The filtrated saturated solution was used for the preparation of further dilution levels by diluting with ISO test water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: STRAUS Clone 5
- Age of parental stock: 2 to 24 hours
- Feeding during test: none

ACCLIMATION
- Acclimation period: at least 2 hours in dilution water
- Acclimation conditions: same as test conditions

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

258 - 265 mg CaCO3/L

Test temperature:

20.2 - 20.6 °C

pH:

7.71 - 7.91

Dissolved oxygen:

8.32 - 8.47 mg/L

Salinity:

NA

Conductivity:

645 - 650 µS/cm

Nominal and measured concentrations:

Nominal concentrations: 0.625, 1.25, 2.50, 5.00, 10.0, 20.0% of the saturated solution
Measured concentrations: 0.0981, 0.256, 0.744, 1.84, 5.84, 11.0 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass flasks
- Type: sealed with screw caps
- Material, size, headspace, fill volume: glass, 4.5 (ID) x 9.5 (H) cm, no headspace, 130 mL capacity
- Aeration: none
- Renewal rate of test solution: The test solutions were renewed after 24 hours. A second set of test vessels were filled with the freshly prepared test solutions and the daphnids were transferred by pipette.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO test water, according to OECD 202, Annex 3
- Culture medium different from test medium: yes, culture medium is Eldent M4, according to OECD 202, Annex 3

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 1340 lux

EFFECT PARAMETERS MEASURED: Immobilisation was determined in all groups after 24 and 48 hours.

RANGE-FINDING STUDY
- Test concentrations: measured concentrations: 2.94, 28.1, 292 µg/L (nominal concentration of the saturated solution - 364 mg/L and dilution levels 1 and 10% of the nominal concentration)
- Results used to determine the conditions for the definitive study: 100% immobilisation in the highest and 80% in the second highest concentrations was observed. Based on these results, a limit concentration of 20% of the saturated solutions was chosen as the highest concentration for the main test.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

10.84 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

9.87 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 11 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no
- Other biological observations: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: yes, adsorption onto the glassware. Adsorption to the glass walls of the test vessels was determined exemplarily for the highest concentration level of 20% of the saturated solution (double determination) in separate replicates without daphnids at the end of both exposure intervals (24 and 48 hours). Adsorption to the glass walls accounted for ≤ 8% of the test item concentration, measured at the start of the respective exposure interval.
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? Yes
A reference test was conducted as an acute immobilisation test (acc. to AQS P 9/2 and OECD 202) in ISO test water (acc. to OECD 202 (2004), Annex 3) under static conditions with a test duration of 24 hours once per month to prove the validity of the test system and test conditions at the test facility. The EC50 - value for the most recent of the monthly performed reference tests was: EC50: 1.95 mg/L (Cl: 1.14 - 3.30 mg/L).

Table 1: Immobilisation Rates after 24 and 48 hours of Exposure in the Definitive Test

               (n = 20, divided into 4 replicates with 5 daphnids each)

Geometric mean measured test item concentrations [µg/L]	IMMOBILISATION [%]
	24 hours						48 hours
	Replicates						Replicates
	1	2		3	4	MV	1	2		3	4	MV
11.0	60	20		20	0	25	60	40		60	80	60
5.84	0	0		0	0	0	0	20		0	0	5
1.84	0	0		0	0	0	20	0		0	20	10
0.744	0	0		0	20	5	20	0		0	20	10
0.256	0	0		0	0	0	0	0		0	0	0
0.0981	0	0		0	0	0	0	0		0	0	0
Control	0	0		0	0	0	0	0		0	0	0

 

Table 2: Measured Concentrations of the Test Item during the Definitive Test

 

Sampling date	2016-04-06 0 hours Start of the exposure interval	2016-04-07 24 hours End of the exposure interval		2016-04-07 24 hours Start of the exposure interval	2016-04-08 48 hours End of the exposure interval		Geometric mean measured test item concentration [µg/L]
Start of analysis	2016-04-06	2016-04-07		2016-04-07	2016-04-08
Dilution level of the saturated solution [%]	Test item
	Meas. conc. [µg/L]	Meas. conc. [µg/L]	%	Meas. conc. [µg/L]	Meas. conc. [µg/L]	%
20.0	16.1^1)	7.41	46	21.3	5.80	27	11.0
10.0	9.94	3.98	40	11.4	2.59	23	5.84
5.00	5.70	0.886	16	4.19	0.547	13	1.84
2.50	3.06	0.327	11	2.49	0.123	5	0.744
1.25	1.47	0.0643	4	1.23	0.0366	3	0.256
0.625	0.634	0.0152	2	0.545	0.0176	3	0.0981
Control	<LOQ	<LOQ		<LOQ	<LOQ

 

Meas. conc. = measured concentration of the test item, dilution factor taken into account

%                 = percentage of the initially measured concentration of the test item

LOQ             = limit of quantification of the analytical method (0.200 µg test item/L)

¹⁾                  = Reanalysed on 2016-04-08

Validity criteria fulfilled:

yes

Conclusions:

Based on the geometric mean measured concentrations of the test item, the 48 hour-EC50 for Daphnia magna was 10.8 µg/L.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202. The study was conducted in a closed system (sealed glass flasks) without headspace under semistatic conditions over a period of 48 hours with five dilution levels of a saturated solution of the test item (nominal concentrations: 0.625 to 20% of the saturated solution) in a geometric series with a separation factor of 2. The validity criteria of the test guideline were fulfilled. The estimated EC50 value (48 hours) of the test item is 10.8 µ.g/L (CI: 6.13 -> 11.0).

Description of key information

The acute immobilisation of the test item to daphnia magna was determined according to OECD 202 (2004). The EC50 value after 48 h is 10.8 µg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

10.8 µg/L

Additional information

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202 (2004). The study was conducted in a closed system (sealed glass flasks) without headspace under semistatic conditions over a period of 48 hours with five dilution levels of a saturated solution of the test item (nominal concentrations: 0.625 to 20% of the saturated solution) in a geometric series with a separation factor of 2. The validity criteria of the test guideline were fulfilled. The estimated EC50 value (48 hours) of the test item is 10.8 µg/L (CI: 6.13 -> 11.0).