N-[3-(isodecyloxy)propyl]propane-1,3-diamine

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

other information

Study period:

2013-04-09 to 2013-04-11

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:

N-[3-(isodecyloxy)propyl]propane-1,3-diamine

EC Number:

276-432-0

EC Name:

N-[3-(isodecyloxy)propyl]propane-1,3-diamine

Cas Number:

72162-46-0

Molecular formula:

C16H36N2O

IUPAC Name:

(3-aminopropyl)({3-[(2-methylnonyl)oxy]propyl})amine

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 2.00 mg/L with dilution water, were tested as follows: 0.0625 - 0.125 - 0.250 - 0.500 - 1.00 - 2.00 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. Sorption to the walls of the test vessel was checked as specified below. For this purpose, additional replicateswithout daphnids were prepared and incubated under test conditions for quantification of sorption to the glassware.
The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11). The sorption of the test item on glass was quantified. Additional replicates of the three highest concentration levels without daphnids were prepared. After sampling for the test media analysis (aqueous phase), the glassware was emptied and an appropriate amount of a suitable solvent was added. The concentration of the test item in this solution was measured and the adsorbed test item amount was calculated from this concentration. Before this procedure, the test vessels were carefully rinsed with demineralised water to remove the remaining test media.
- Sample storage conditions before analysis: All original samples were stored at room temperature. Prepared samples were stored at room temperature until analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution (2.00 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 treated with ultrasound for 5 minutes at room temperature and stirred thereafter with approximately 1100 rpm for 2 h at room temperature.



- Eluate: Water of the river Innerste was used. This river is located near 31157 Ruthe-Sarstedt, Germany (Latitude 52.25°, Longitude 9.83°). The natural river water was stored at 15 ± 2 °C until use in the test.

Water Parameters of the River Water
River Innerste
Location Amtshof K514, km 26,
31157 Ruthe-Sarstedt, Germany
Latitude:52.25°, Longitude: 9.83°
Sampling Date 2012-09-17
Weather on Day of Sampling Sunny, slightly clouded;
approx. 18 °C
Colour Yellowish to brownish, clear
pH 7.56
Conductivity [µS/cm] 898
Dissolved Oxygen [mg O2/L] 8.83
NPOC [mg C/L] 3.13
Ammonium-N [mg N/L] 0.05
Nitrate-N [mg N/L] 1.70
Total Nitrogen [mg N/L] 2.70
o-Phosphate-p [mg P/L] < 0.20
Total Phosphate [mg P/L] < 0.20
Suspended Matter [mg/L] 16.4
Total Hardness [mg CO3/L] 283
Storage conditions 15 ± 2°C


- Differential loading: 0.0625 - 0.125 - 0.250 - 0.500 - 1.00 - 2.00 mg/L.
- Control: Natural river 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:
Water of the river Innerste was used. This river is located near 31157 Ruthe-Sarstedt, Germany (Latitude 52.25°, Longitude 9.83°). The natural river water was stored at 15 ± 2 °C until use in the test.


Water Parameters of the River Water
River Innerste
Location Amtshof K514, km 26,
31157 Ruthe-Sarstedt, Germany
Latitude:52.25°, Longitude: 9.83°
Sampling Date 2012-09-17
Weather on Day of Sampling Sunny, slightly clouded;
approx. 18 °C
Colour Yellowish to brownish, clear
pH 7.56
Conductivity [µS/cm] 898
Dissolved Oxygen [mg O2/L] 8.83
NPOC [mg C/L] 3.13
Ammonium-N [mg N/L] 0.05
Nitrate-N [mg N/L] 1.70
Total Nitrogen [mg N/L] 2.70
o-Phosphate-p [mg P/L] < 0.20
Total Phosphate [mg P/L] < 0.20
Suspended Matter [mg/L] 16.4
Total Hardness [mg CO3/L] 283
Storage conditions 15 ± 2°C

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:

Natural river water 0 h: 304 (mg CaCO3/L)
Natural river water 24 h: 255 (mg CaCO3/L)

Test temperature:

0 h: 19.1 °C
24 h: 19.3 °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)
Nominal concentration
of the test item
[mg/L] pH-Value
2.00 7.83
1.00 7.84
0.500 7.80
0.250 7.78
0.125 7.78
0.0625 7.81
Control 7.70

Water Quality Parameters at the End of the 1st Exposure Interval (24 h)
(measured in all replicates)
Nominal concentration
of the test item
[µg/L] pH-Values
Replicates
1 2 3 4
2.00 8.19 8.20 8.20 8.20
1.00 8.20 8.21 8.21 8.20
0.500 8.21 8.20 8.20 8.18
0.250 8.21 8.20 8.20 8.21
0.125 8.19 8.19 8.20 8.20
0.0625 8.09 8.16 8.18 8.20
Control 8.30 8.23 8.21 8.20

Water Quality Parameters at the Start of the 2nd Exposure Interval (24 h)
(measured in one additional replicate per concentration level and control)
Nominal concentration
of the test item
[µg/L] pH-Value
2.00 Not determined, due to 100 % mortality
1.00 Not determined, due to 100 % mortality
0.500 7.94
0.250 7.91
0.125 7.91
0.0625 7.94
Control 7.77

Water Quality Parameters at the End of the 2nd Exposure Interval (48 h)
(measured in all replicates)
Nominal concentration
of the test item
[µg/L] pH-Values
Replicates
1 2 3 4
2.00 Not determined, due to 100 % mortality
1.00 Not determined, due to 100 % mortality
0.500 8.23 8.23 8.23 8.22
0.250 8.17 8.20 8.20 8.21
0.125 8.18 8.17 8.18 8.18
0.0625 8.12 8.19 8.18 8.16
Control 8.04 8.15 8.19 8.19

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)
Nominal concentration
of the test item
[µg/L] Dissolved O2-Concentration
2.00 8.43
1.00 8.71
0.500 8.87
0.250 9.00
0.125 9.05
0.0625 9.10
Control 9.40

Water Quality Parameters at the End of the 1st Exposure Interval (24 h)
(measured in all replicates)
Nominal concentration
of the test item
[µg/L] Dissolved O2-Concentration
Replicates
1 2 3 4
2.00 8.27 8.28 8.25 8.27
1.00 8.28 8.30 8.29 8.29
0.500 8.28 8.29 8.30 8.26
0.250 8.26 8.31 8.33 8.30
0.125 8.27 8.30 8.35 8.27
0.0625 8.37 8.30 8.24 8.27
Control 8.17 8.06 8.08 8.02

Water Quality Parameters at the Start of the 2nd Exposure Interval (24 h)
(measured in one additional replicate per concentration level and control)
Nominal concentration
of the test item
[µg/L] Dissolved O2-Concentration
2.00 Not determined, due to 100 % mortality
1.00 Not determined, due to 100 % mortality
0.500 8.90
0.250 8.98
0.125 9.00
0.0625 8.81
Control 9.45

Water Quality Parameters at the End of the 2nd Exposure Interval (48 h)
(measured in all replicates)
Nominal concentration
of the test item
[µg/L] Dissolved O2-Concentration
Replicates
1 2 3 4
2.00 Not determined, due to 100 % mortality
1.00 Not determined, due to 100 % mortality
0.500 7.86 7.69 7.59 7.56
0.250 7.69 7.28 7.51 7.51
0.125 7.94 7.80 7.91 7.70
0.0625 7.86 7.93 7.85 7.65
Control 7.74 7.77 7.81 7.84

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), 50 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:1106 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
5.00 100 100 100 100 100 100
0.500 0 0 0 60 30 45
0.0500 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

0.221 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: (0.128 - 0.406)

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: 2.14 (CI 1.92 - 2.34) 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)

Nominal test item concentration [mg/L]	IMMOBILISATION [%]
	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
2.00	100	100	100	100	100	100	100	100	100	100
1.00	100	100	100	100	100	100	100	100	100	100
0.500	0	40	40	20	25	100	100	100	100	100
0.250	0	0	0	0	0	80	80	100	100	90
0.125	0	0	0	0	0	0	0	0	0	0
0.0625	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 nominal concentrations of the test item N-[3-(Isodecyloxy)propyl]propane-1,3-diamine the 48 h-EC50 for Daphnia magna was 0.235 mg/L (95 % confidence limits: 0.133 – 0.413 mg/L).
The NOEC after 48 h was 0.125 mg/L. The LOEC after 48 h was 0.250 mg/L.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item        N-[3-(Isodecyloxy)propyl]propane-1,3-diamine(batch number: F-212290) were determined according to OECD 202 (2004) from 2013-04-09 to 2013-04-11 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 N-[3-(Isodecyloxy)propyl]propane-1,3-diaminein the range of 0.0625 to 2.00 mg/L, prepared with natural river water in a geometric series with a separation factor of 2. Twenty daphnids were exposed to each concentration level and the control.

The concentrations of the test item N-[3-(Isodecyloxy)propyl]propane-1,3-diaminewere 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 freshly prepared stock solutions of 2.00 mg/L were analysed at the start of the exposure phase (0 h) and at the water renewal (24 h).

The measured concentrations of the test item at the start of the exposure intervals (0 and 24 h) were in the range of 97 to 116 % 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 101 to 110 % of the nominal values. The recovery rates of the stock solutions of 2.00 mg/L were 100 % (0 h) and 110 % (24 h) of the nominal value.

Adsorption to the glass walls of the test vessels was determined exemplarily at the three highest concentration levels in separate replicates without daphnids. Due to 100 % mortality at the concentration levels of 1.00 and 2.00 mg/L after 24 h, adsorption was determined after 48 h at the concentration levels of 0.125 and 0.250 mg/L instead. Adsorption to the glass walls was ≤ 1 % of the nominal test item concentrations at the concentration levels of 0.125 to 2.00 mg/L.

The test item N-[3-(Isodecyloxy)propyl]propane-1,3-diamine has a low water solubility and sorbs to organic and inorganic materials by different mechanisms. The sorption processes are mostly non-linear, means are concentration dependent. Due to these properties the test item is difficult to test in synthetic water (e.g. sorption to the test organisms and glass walls of the test vessels) and results from such tests depend from the test settings applied. Using natural river water which contains particulate as well as dissolved organic carbon to which the test item can sorb partially reduces the difficulties encountered in tests with synthetic water (e.g. preventing that the test item settles onto surfaces). The sorbed fraction of the test item is often difficult to extract from the test system which normally leads to low analytical recoveries. The recoveries of the test item show, that the test concentrations were successfully maintained for the duration of the test. Adsorption to the glass walls was negligible. Therefore, the nominal concentrations were used for assessment of the endpoints instead of measured ones.

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 nominal concentrations of N-[3-(Isodecyloxy)propyl]propane-1,3-diamine)

Endpoints	Test duration [h]	N-[3-(Isodecyloxy)propyl]propane-1,3-diamine
		Nominal test item concentrations [mg/L]	95 % confidence limits [mg/L]
EC10	24	0.483	0.267 – 0.933
	48	0.221	0.128 – 0.406
EC50	24	0.517	0.282 – 0.949
	48	0.235	0.133 – 0.413
EC100	24	1.00	Not applicable
	48	0.500	Not applicable
NOEC	48	0.125	Not applicable
LOEC	48	0.250	Not applicable