N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20±2°C

pH:

8.1 to 8.3

Dissolved oxygen:

≥ 8.0 mg/l (88% of saturation).

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.073 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.025 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

0.13 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

- Toxicity of the test material to daphnids followed a positive dose response for non-mortality/immobility.
- Of the validity criteria for this study, only the requirement for test concentration to be ≥ 80% of nominal was unfulfilled. The test material stability appeared to be concentration dependent. However, this was not considered to affect the validity of the study as mean-measured concentrations were used to determine effect concentrations. Effects were also only noted at test concentrations where measured test concentrations were within the range of ≥ 80% of nominal and hence not as greatly affected by the concentration dependant instability. The results are considered to be reliable under the conditions of the study.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the measured 48h EC50 and NOEC were determined to be 0.073 and 0.05 mg/L, respectively.

Executive summary:

A study was conducted to determine the toxicity of the test substance to water flea (Daphnia magna) according to EPA OPPTS Guideline 850.1010, in compliance with GLP. For each test concentration, 20 Daphnia were exposed to the test substance for 48 h in a static test system. After 24 and 48 h, the immobilised Daphnia were counted. Analytical dose verification was conducted. Toxicity of the test substance followed a positive dose response for mortality/immobility. Under the study conditions, the measured 48h EC50 and NOEC were determined to be 0.073 and 0.05 mg/L, respectively (Drottar, 2000).

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 13, 2002 to September 26, 2002

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)

Deviations:

yes

Remarks:

see 'Principles of method if other than guideline'

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

see 'Principles of method if other than guideline'

Principles of method if other than guideline:

The study was conducted according to the OECD Guideline 202 and EU Method C.2, in compliance with GLP. The only deviation was that analytical quantification of the applied test concentrations was not possible due to the absence of a suitable analytical method. Therefore only the stock solution used to prepare the test concentrations were analyzed. However, based on the high water solubility and stability of the test substance it was considered reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Triamine content: 85.3%; Total amine content (commercial grade): 100%

Analytical monitoring:

yes

Details on sampling:

Although quantification of the applied concentrations was not possible due to the absence of a sensitive analytical method, the stock solution used to prepare the test concentrations was analyzed. These analyses showed that the stock solution was stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.

Vehicle:

no

Details on test solutions:

The test substance was soluble in water.
A stock solution of approximately 0.1 g/L of test substance was prepared as follows:
- To an accurately measured amount of 0.1127 g of test substance approximately 120 mL of deionized water of approximately 50°C was added. The test substance dissolved well under stirring. After cooling of deionized water was added up to a final volume of 1000 mL. A homogeneous, clear stock solution was obtained. It was kept for stirring at room temperature before being used for the preparation of the test solutions at the start of the test.
- The test solutions were prepared by addition of the required amounts of stock solution to the test medium to obtain the following concentrations: 0.01, 0.02, 0.04, 0.08 and 0.16 mg/L. A control containing only test medium was included in the test.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISMS:
- Strain: Daphnia magna (water fleas)
- Source/supplier: Aquasense, Amsterdam
- Breeding method: According to OECD 202
- Age: < 24 h and were obtained from parent animals having an age of 2-4 weeks
- Feeding: Algae; chlorella vulgaris
- Pretreatment: daily 0.1-0.2 C/Daphnia
- Feeding during test: No

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

12°dH

Test temperature:

The temperature in the test vessels during the test period ranged from 20.6 to 21.4°C.

pH:

The maximum variation of pH observed during the test was 0.2 units.

Dissolved oxygen:

The maximum variation in oxygen concentration during the test was 0.3 mg/L

Salinity:

As test medium so called Dutch Standard water (DSW) was used, with a pH of approximately 8.2 and a hardness of approximately 12°dH, containing per liter of deionized water: 100 mg of NaHCO3, 20 mg of KHCO3, 200 mg of CaCl2•2H2O and 180 mg of MgSO4•7H2O. The test medium was aerated before being used in the test. The air was water-saturated and purified by an active coal and cotton filter.

Nominal and measured concentrations:

-Nominal concentrations: 0.01, 0.02, 0.04, 0.08 and 0.16 mg/L.
- The available analytical method was not suitable to quantify concentrations below 10 mg/L. Therefore only the stock solutions used to prepare the test concentrations were analyzed. The measured concentration of the undiluted stock solution (109.8 mg/L) differs only slightly from the nominal concentration of the stock solution (112.7 mg/L). These results show that the stock solution was stable even after storage for almost 7 months in the refrigerator. All concentrations mentioned in this report are therefore based on nominal concentrations. Considering the high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.

Details on test conditions:

TEST SYSTEM
- Test type: Static
- Renewal of test solution: No
- Exposure vessel type: 400 mL glass beakers containing 250 mL test solution
- Number of replicates, individuals per replicate: 4 vessels at each concentration and the control with 5 Daphnia per vessel
- Control group: Yes, concurrent vehicle
- Vehicle, solvent: deionized water
- Concentration of vehicle/ solvent: 0.1 g/L
- Aeration: No
- Other: The daphnids were randomly placed in the test solutions and the test vessels were positioned in a random manner. During the test the vessels were covered with glass plates. The test solutions were not aerated. The animals were not fed during the test.

- TEST MEDIUM / WATER PARAMETERS
- Dilution water source: Dutch Standard Water; Containing per liter of dionized water 100 mg Na HCO3, 20 mg KHCO3, 200 mg CaCl2.2H2O and 180 mg MgSO4.7H2O
- Hardness: 12°dH
- pH: 7.9-8.2
- Oxygen content: 8.7-9.1 mg/L
- Holding water: DSW TEST SYSTEM
- Concentrations: 0, 0.01, 0.02, 0.04, 0.08, 0.16 mg/L
- Test temperature: 20.6-21.4 °C
- Dissolved oxygen: 8.7-9.1 mg/L
- pH: 7.9-8.2
- Adjustment of pH: No
- Intensity of irradiation: Ambient light provided by fluorescent tubes
- Photoperiod: 16 h photoperiod daily

DURATION OF THE TEST: 48 h

TEST PARAMETER: The daphnids were considered immobile when they were not able to swim for 15 seconds after gentile agitation of the test vessel. Also sub-lethal effects such as floating at the surface were recorded.
- Observation interval: Immobility and sub-lethal effects were recorded at approximately 24 and 48 h.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.077 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Based on total amine content

Basis for effect:

mobility

Remarks on result:

other: (0.066 - 0.092)

Key result

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

0.04 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Based on total amine content

Basis for effect:

mobility

Remarks on result:

other: Considering the high water solubility and stability of the test substance, the actual concentrations were considered identical to the nominal concentrations.

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

0.16 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Based on total amine content

Basis for effect:

mobility

Remarks on result:

other: Considering the high water solubility and stability of the test substance, the actual concentrations were considered identical to the nominal concentrations.

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.04 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Based on total amine content

Basis for effect:

mobility

Remarks on result:

other: Considering the high water solubility and stability of the test substance, the actual concentrations were considered identical to the nominal concentrations.

Details on results:

The chemical analyses of the stocksolutions showed that the stock solution was stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.

Results with reference substance (positive control):

-

Reported statistics and error estimates:

The EC50 was calculated with the computer program TOXCALC version 5.0 using the trimmed Spearman-Kärber method. The concentrations causing zero and 100% immobility was derived directly from the test observations.

RESULTS: EXPOSED
- Nominal/measured concentrations: 0, 0.01, 0.02, 0.04, 0.08, 0.16 mg/L (nominal)
- Effect data (Immobilisation): 
          Immobile Daphnia
conc mg/L       24 h                 48 h 
0	  0	 1
0.01	  0	 2
0.02	  1	 2
0.04	  0	 1
0.08	  0	 11
0.16	  9	 20

The EC50 - 48 h was calculated to be 0.078 mg/L with 95 % confidence limits of 0.066 and 0.092 mg/L. At 24 h no effects were too low to derive an EC50. The NOEC 48 h amounted to 0.04 mg/L, the LOEC 48 h was 0.08 mg/L. The
EC100 was 0.16 mg/L and was observed after 48 h. A dose-response related increase in mortality was observed.

- Other effects: Sublethal effects were not observed in the test. VALIDITY CRITERIA: Based on the following data, the validity criteria of the test was considered as fulfilled: - immobilisation of control animals was <10%, - the control animals were not staying at the surface, - the concentration of dissolved oxygen in all test vessels was >3 mg/L and - the concentration of test substance >= 80% of initial concentration during test.

Table 1

Test results

 

Test substance	Batches	Number of mobile animals
(mg/L)		0 hours	24 hours	48 hours
Control	Batch I Batch II Batch III Batch IV Total	5 5 5 5 20	5 5 5 5 20	5 4 5 5 19
0.01	Batch I Batch II Batch III Batch IV Total	5 5 5 5 20	5 5 5 5 20	4 5 5 4 18
0.02	Batch I Batch II Batch III Batch IV Total	5 5 5 5 20	4 5 5 5 19	4 4 5 5 18
0.04	Batch I Batch II Batch III Batch IV Total	5 5 5 5 20	5 5 5 5 20	5 5 4 5 19
0.08	Batch I Batch II Batch III Batch IV Total	5 5 5 5 20	5 5 5 5 20	1 2 1 5 9
0.16	Batch I Batch II Batch III Batch IV Total	5 5 5 5 20	3 1 3 4 11	0 0 0 0 0

 

Table 2

Statistical evaluation

 

Time (hours)	EC50(mg/l)	95% confidence limits (mg/l)
24	n.d	n.d	-	n.d
48	0.0775	0.0655	-	0.0918

n.d = not determined; could not be calculated

 

Table 3

Oxygen measurements

Nominal concentration (mg/l)	0	48 h
Control	8.9	8.9
0.01	8.8	9.1
0.02	8.8	9.1
0.04	8.7	8.6
0.08	8.7	8.9
0.16	8.7	8.9

 

 

Table 4

pH-measurements

Nominal concentration (mg/L)	0	48 h
Control	8.1	7.9
0.01	8.1	7.9
0.02	8.1	8.0
0.04	8.1	8.0
0.08	8.1	8.0
0.16	8.2	8.0

 

Table 5

Results of the chemical analyses

 

	Measured Concentration in 10 times diluted stock solution (mg/L)	Average measured concentration (mg/L)	Standard deviation
First analyses stock 13022002	109.86	109.80	0.08
Second analyses 13022002	109.74

 

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the nominal 48 h EC50 and NOEC of the test substance were calculated to be 0.078 and 0.04 mg/L, respectively. The test substance was found to be toxic to Daphnia magna after 48 h at concentration levels of 0.08 mg/L and higher.

Executive summary:

A study was conducted to determine the acute toxicity of the test substance to water flea (Daphnia magna) according to the OECD Guideline 202 and EU Method C.2, in compliance with GLP. In this study, 4 replicates of 5 juvenile Daphnia per concentration were exposed to 0.01, 0.02, 0.04, 0.08 and 0.16 mg/L nominal test concentrations under static conditions for 48 h. Immobility and sub-lethal effects were recorded at 24 and 48 h. Although quantification of the applied concentrations was not possible due to the absence of a suitable analytical method, it was possible to analyze the two stock solutions used to prepare the test concentrations. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility and stability of the test substance, it was considered reasonable to assume that the actual concentrations would be identical to the nominal concentrations during the entire test. All the validity criteria were considered to be fulfilled. Under the study conditions, the nominal 48 h EC50 and NOEC of the test substance were calculated to be 0.078 and 0.04 mg/L, respectively. The test substance was found to be toxic to Daphnia magna after 48 h at concentration levels of 0.08 mg/L and higher (Geurts, 2002).

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1989

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: The analytical verification of the test concentration are partially reliable. Further, only a 24 hours exposure was investigated.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

0.67 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

0.095 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the measured 24h EC50 and NOEC values in Daphnia magna were determined to be 0.67 and 0.095 mg/L, respectively. However, the results should be considered with caution as the analytical verification of the test concentration were only partially reliable. Also, only a 24 h exposure was investigated.

Executive summary:

A study was conducted to determine toxicity of the test substance to water flea (Daphnia magna) according to OECD Guideline 202, in compliance with GLP. For each test concentration, 20 Daphnia were exposed for 24 h in a static test system. After 24 h, immobilised Daphnia were counted. Toxicity of the test substance to Daphnids followed a positive dose response for non-mortality/immobility. Under the study conditions, the measured 24h EC50 and NOEC values in Daphnia magna were determined to be 0.67 and 0.095 mg/L, respectively. However, the results should be considered with caution as the analytical verification of the test concentration were only partially reliable. Also, only a 24 h exposure was investigated (Ritter, 1989).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.078 mg/L

Additional information

A study was conducted to determine the acute toxicity of the test substance to water flea (Daphnia magna) according to the OECD Guideline 202 and EU Method C.2, in compliance with GLP. In this study, 4 replicates of 5 juvenile Daphnia per concentration were exposed to 0.01, 0.02, 0.04, 0.08 and 0.16 mg/L nominal test concentrations under static conditions for 48 h. Immobility and sub-lethal effects were recorded at 24 and 48 h. Although quantification of the applied concentrations was not possible due to the absence of a suitable analytical method, it was possible to analyze the two stock solutions used to prepare the test concentrations. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility and stability of the test substance, it was considered reasonable to assume that the actual concentrations would be identical to the nominal concentrations during the entire test. All the validity criteria were considered to be fulfilled. Under the study conditions, the nominal 48 h EC50 and NOEC of the test substance were calculated to be 0.078 and 0.04 mg/L, respectively. The test substance was found to be toxic to Daphnia magna after 48 h at concentration levels of 0.08 mg/L and higher (Geurts, 2002).

A study was conducted to determine the toxicity of the test substance to water flea (Daphnia magna) according to EPA OPPTS Guideline 850.1010, in compliance with GLP. For each test concentration, 20 Daphnia were exposed to the test substance for 48 h in a static test system. After 24 and 48 h, the immobilised Daphnia were counted. Analytical dose verification was conducted. Toxicity of the test substance followed a positive dose response for mortality/immobility. Under the study conditions, the measured 48h EC50 and NOEC were determined to be 0.073 and 0.05 mg/L, respectively (Drottar, 2000).

A study was conducted to determine toxicity of the test substance to water flea (Daphnia magna) according to OECD Guideline 202, in compliance with GLP. For each test concentration, 20 Daphnia were exposed for 24 h in a static test system. After 24 h, immobilised Daphnia were counted. Toxicity of the test substance to Daphnids followed a positive dose response for non-mortality/immobility. Under the study conditions, the measured 24h EC50 and NOEC values in Daphnia magna were determined to be 0.67 and 0.095 mg/L, respectively. However, the results should be considered with caution as the analytical verification of the test concentration were only partially reliable. Also, only a 24 h exposure was investigated (Ritter, 1989).