3-(isodecyloxy)propiononitrile

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:

2013-04-03 to 2013-05-24

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-(isodecyloxy)propiononitrile

EC Number:

264-840-1

EC Name:

3-(isodecyloxy)propiononitrile

Cas Number:

64354-92-3

Molecular formula:

C13H25NO

IUPAC Name:

3-(decylalkyl-(branched)oxy)-propiononitrile

Details on test material:

- Name of test material (as cited in study report): 3-(Isodecyloxy)propionitrile
- Physical state: Slightly yellowish, clear liquid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:6 test item concentrations in a geometric series with a separation factor of 2, prepared by diluting the stock solution of 3.00 mg/L with dilution water, were tested as follows: 0.938 - 0.188 - 0.375 - 0.750 - 1.50 - 3.00 mg/L.


- Sampling method: All concentration levels and the control were analytically verified by LC-MS/MS at the start (0 and 24 h) and at the end of both exposure intervals (24 and 48 h). At the start of the exposure intervals (0 and 24 h), sampling was carried out immediately after preparation of the test concentrations. At the end of the exposure intervals (24 and 48 h), samples were taken directly from the test vessels.
The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All original and diluted samples were stored at room temperature until sample preparation and until analysis, if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Prior to weighing, the test item was treated with ultrasound for 5 minutes at 40 °C. The stock solution (3.00 mg/L of the test item were weighed out) was freshly prepared with dilution water before the start of both exposure intervals (0 and 24 h). The stock solution was mixed with an ultraturrax (19600 rpm for 1 minute) and stirred with a magnetic stirrer with 1100 rpm for 2 h at room temperature.


- Eluate: Dilution water

- Differential loading:nominal 0.0938 - 0.188 - 0.375 - 0.750 - 1.50 - 3.00 mg/L.
corresponding to geometric mean measured concentrations of: 0.0727 - 0.146 - 0.278 - 0.564 - 1.20 - 2.50 mg/L
- Control: Dilution water without test item tested under the same conditions as the test groups.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

DILUTION WATER:
Dilution water ISO test water, according to OECD 202, Annex 3:
Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2  2 H2O 294
MgSO4  7 H2O 123
pH-value: 7.8  0.2

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Test conditions

Hardness:

Dilution water 0 h: 231 (mg CaCO3/L)
Dilution water 24 h: 265 (mg CaCO3/L)

Test temperature:

0 h: 21.8 °C
24 h: 20.8 °C

pH:

Water Quality Parameters at the Start of the 1st Exposure Interval (0 h)
(measured in one additional replicate per concentration level and control)
Geometric mean
measured test
item concentrations
[mg/L] pH-Value
2.50 7.85
1.20 7.88
0.564 7.88
0.278 7.90
0.146 7.90
0.0727 7.89
Control 7.96

Water Quality Parameters at the End of the 1st Exposure Interval (24 h)
(measured in all replicates)
Geometric mean
measured test
item concentrations
[mg/L] pH-Values
Replicates
1 2 3 4
2.50 7.69 7.68 7.68 7.68
1.20 7.66 7.66 7.67 7.67
0.564 7.68 7.68 7.67 7.66
0.278 7.67 7.67 7.68 7.68
0.146 7.67 7.63 7.67 7.67
0.0727 7.67 7.66 7.67 7.67
Control 7.63 7.65 7.65 7.68

Water Quality Parameters at the Start of the 2nd Exposure Interval (24 h)
(measured in one additional replicate per concentration level and control)
Geometric mean
measured test
item concentrations
[mg/L] pH-Value
2.50 7.73
1.20 7.73
0.564 7.73
0.278 7.72
0.146 7.70
0.0727 7.62
Control 7.84

Water Quality Parameters at the End of the 2nd Exposure Interval (48 h)
(measured in all replicates)
Geometric mean
measured test
item concentrations
[mg/L] pH-Values
Replicates
1 2 3 4
2.50 7.70 7.71 n.d. 7.71
1.20 7.70 7.70 7.70 7.69
0.564 7.69 7.69 7.68 7.69
0.278 7.68 7.68 7.68 7.68
0.146 7.68 7.67 7.69 7.69
0.0727 7.66 7.67 7.68 7.67
Control 7.76 7.73 7.72 7.72

Dissolved oxygen:

Water Quality Parameters at the Start of the 1st Exposure Interval (0 h)
(measured in one additional replicate per concentration level and control)
Geometric mean
measured test
item concentrations
[mg/L] Dissolved O2-Concentration
2.50 8.16
1.20 8.26
0.564 8.32
0.278 8.34
0.146 8.33
0.0727 8.37
Control 8.36

Water Quality Parameters at the End of the 1st Exposure Interval (24 h)
(measured in all replicates)
Geometric mean
measured test
item concentrations
[mg/L] Dissolved O2-Concentration
Replicates
1 2 3 4
2.50 8.11 8.12 8.11 8.04
1.20 8.07 8.14 8.11 8.10
0.564 8.19 8.17 8.14 8.14
0.278 8.21 8.23 8.25 8.26
0.146 8.25 8.25 8.19 8.25
0.0727 8.20 8.23 8.24 8.24
Control 8.20 8.18 8.26 8.62

Water Quality Parameters at the Start of the 2nd Exposure Interval (24 h)
(measured in one additional replicate per concentration level and control)
Geometric mean
measured test
item concentrations
[mg/L] Dissolved O2-Concentration
2.50 8.20
1.20 8.21
0.564 8.39
0.278 8.49
0.146 8.53
0.0727 8.47
Control 8.72

Water Quality Parameters at the End of the 2nd Exposure Interval (48 h)
(measured in all replicates)
Geometric mean
measured test
item concentrations
[mg/L] Dissolved O2-Concentration
Replicates
1 2 3 4
2.50 8.17 8.22 n.d. 8.04
1.20 7.90 7.92 8.08 8.11
0.564 8.03 8.05 8.26 8.27
0.278 8.15 8.19 8.21 8.18
0.146 8.17 8.18 8.16 8.19
0.0727 8.33 8.25 8.23 8.19
Control 8.03 8.16 8.29 8.30
n.d. = not determined, due to 100 % mortality

Salinity:

Not measured, freshwater

Nominal and measured concentrations:

please refer to "Any other information on materials and methods"

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 20 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): semi-static test with a renewal of the test solutions after 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start:617 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start, at water renewal after 24 h and test end


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilisation Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal test item
concentration
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
10.0 80 90 85 100 100 100
1.00 0 0 0 50 40 45
0.100 0 0 0 10 0 5
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

1.09 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: (0.565 - 2.24)

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.17 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: (0.596-2.28)

Details on results:

- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.80 (CI 1.52 - 2.22) mg/L

Reported statistics and error estimates:

EC100-values after 24 and 48 h and the NOEC and LOEC after 48 h were deduced directly from the observation data, without mathematical calculation. EC10- and EC50-values were calculated by sigmoidal dose-response regression. The confidence intervals were calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism5. The concentration effect relationships are shown graphically. The EC50-value for the reference item and its confidence limits were calculated accordingly.

Any other information on results incl. tables

Immobilisation Rates after 24 and 48 h of Exposure in the Definitive Test

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

3-(Isodecyloxy)propionitrile		IMMOBILISATION [%]
Nominal test item concentration [mg/L]	Geometric mean measured test item concentration [mg/L]	24 h					48 h
		Replicates					Replicates
		1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
3.00	2.50	80	60	100	40	70	100	100	100	100	100
1.50	1.20	20	20	20	20	20	80	100	60	40	70
0.750	0.564	0	0	0	0	0	0	0	0	0	0
0.375	0.278	0	0	0	0	0	0	0	0	0	0
0.188	0.146	0	0	0	0	0	0	0	0	0	0
0.0938	0.0727	0	0	0	0	0	0	0	0	0	0
Control		0	0	0	0	0	0	0	0	0	0

MV       = mean value

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the geometric mean measured concentrations of the test item 3-(Isodecyloxy)propionitrile the 48 h-EC50 for Daphnia magna was 1.17 mg/L (95 % confidence limits: 0.596 – 2.28 mg/L).
The NOEC after 48 h was 0.564 mg/L. The LOEC after 48 h was 1.20 mg/L.

Based on the nominal concentrations of the test item 3-(Isodecyloxy)propionitrile the 48 h-EC50 for Daphnia magna was 1.46 mg/L (95 % confidence limits: 0.787 – 2.72 mg/L).
The NOEC after 48 h was 0.750 mg/L. The LOEC after 48 h was 1.50 mg/L.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item 3 -(Isodecyloxy)proionitrile (batch number: F-212252) were determined according to OECD 202 (2004) from 2013-04-03 to 2013 -05 -24, with the definitive exposure phase from 2013 -05 -22 to 2013 -05 -24, at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str.1, 31157 Sarstedt, Germany.

The study was conducted under semi-static conditions over a period of 48 h with 6concentrations of the test item 3 -(Isodecyloxy)propionitrile in the range of 0.0938 to 3.00 mg/L, prepared with dilution water in a geometric series with a separation factor of 2.

The test item 3 -(Isodecyloxy)propionitrile has a water solubility of approximately 3 mg/L. Therefore, the test concentrations were selected not exceeding the solubilitx limit. Twenty daphnids were exposed to each concentration level and the control.

The concentrations of the test item 3 -(Isodecyloxy)propionitrile were analytically verified by LC-MS/MS at the start of the exposure intervals (0 and 24 h) and at the end of the exposure intervals (24 and 48 h) in all concentration levels and the control.

The measured concentrations of the test item at the start of the exposure intervals (0 and 24 h) were in the range of 83 to 107 % of the nominal values. At the end of the exposure intervals (24 and 48 h), the measured concentrations of the test item were in the range of 56 to 77 % of the nominal values. The geomtric mean measured concentrations of the test item were calculated to be: 0.0727 - 0.146 - 0.278 - 0.564 - 1.20 - 2.50 mg/L.

The effect values given are based on the geometric mean measured concentrations of the test item 3 -(Isodecyloxy)propionitrile and additionally based on the nominal test item concentrations.

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 and 24 h) and at the end of the exposure intervals (24 and 48 h), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

EC₁₀-, EC₅₀- (with Confidence Interval), EC₁₀₀-Values, NOEC and LOEC

(based on the geometric mean measured and the nominal concentrations of the test item 3 -(Isodecyloxy)propionitrile)

Endpoints	Test duration [h]	3 -(Isodecyloxy)propionitrile
		Geometric mean measured test item concentrations [mg/L]	Nominal test item concentrations [mg/L]
EC10 (with 95 % confidence limits)	24	0.940 (CI: 0.179 – 1.21)	1.19 (CI: 0.928 – 1.51)
	48	1.09 (CI: 0.565 – 2.24)	1.37 (CI: 0.750 – 2.66
EC50 (with 95 % confidence limits)	24	1.89 (CI: 1.66 – 2.16)	2.31 (CI: 2.04 – 2.61)
	48	1.17 (CI: 0.596 – 2.28)	1.46 (CI: 0.787 – 2.72)
EC100	24	> 2.50 (Not applicable)	> 3.00 (Not applicable)
	48	2.50 (Not applicable)	3.00 (Not applicable)
NOEC	48	0.564 (Not applicable)	0.750 (Not applicable)
LOEC	48	1.20 (Not applicable)	1.50 (Not applicable)