N-[3-(dimethylamino)propyl]stearamide monoacetate

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:

2014-11-10 to 2015-03-23

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

This study followed the principles of OECD Guideline 203 (1992), but used a non-standard species due to the jurisdiction in which the study was conducted. ECHA Guidance R.17b (2017) indicates "Non-standard species can also be accepted. However, these should be properly identified and characterised in order to ensure that the test method is suitable." Gobiocypris rarus is a species of cyprinid fish endemic to China, and is a standard test species for this jurisdiction. The study is therefore considered suitable for aquatic hazard assessment under REACH.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

yes

Remarks:

Non-standard species used Gobiocypris rarus.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Samples were taken at 0, 24, 48, 72 and 96 h in duplicates or more. One sample was analysed straight after pre-treatmeant, the other was stored

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

The test solution was directly added to water and then stirred for 30 min. Test solutions were colourless and clear.

Test organisms (species):

other: Rare minnow (Gobiocypris rarus)

Details on test organisms:

TEST ORGANISM
- Common name: Rare minnow
- Strain: Not specified
- Source: Institute of Hydrobiology, Chinese Academy of Sciences (Batch No. F20141015)
- Age at study initiation: the fish were acquired as fry and then held for 40 days before definite test.
- Length at study initiation (length definition, mean, range and SD):2.56 cm(SD: 6.24%, range not specified)
- Weight at study initiation (mean and range, SD): 0.259 g (SD: 5.47%, range not specified)
- Method of breeding: Not specified
- Maintenance of the brood fish: The fish were fed daily, they were held without food for approx. 24 hours before testing.

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): Same as test conditions
- Type and amount of food during acclimation: The food ingredients were: Crude protein >36%, Crude Fat >2 %, Crude Fibre <3 %, Crude Ash <12%, Moisture < 10%.
- Feeding frequency during acclimation: The fish were fed daily and then were starved for 24 hours before the test.- Health during acclimation (any mortality observed):After the initial 48 hour settling in period, no mortality was observed.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

126- 135 mg CaCO3/L

Test temperature:

23.0 - 23.2 °C

pH:

7.67 - 7.92

Dissolved oxygen:

between 73 and 92% of air saturation

Salinity:

Not relevant

Nominal and measured concentrations:

The nominal concentrations were: 1.0, 2.0, 4.0, 6.0, 8.0, 10.0 mg/L, the measured concentrations averaged at 0.805, 1.84, 3.94, 5.82, 7.80 and 9.86, respectively, therefore all falling within the acceptable range of -/+ 20% of the nominal concentration.

Details on test conditions:

TEST SYSTEM
- Test vessel: tank
- Material, size, headspace, fill volume: 3 L of test solution in each vessel.
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): NA
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: tap water that was dechlorinated for at least 24 h was used
- Total organic carbon:1.3 mg/L
- Particulate matter:<0.001 mg/L
- Metals: All (As, Cd, Pb, Hg, Co, Ni, Cu, Cr, Fe, Zn, Ag) below acceptable value (OECD 203)
- Pesticides:NS
- Chlorine: NS
- Alkalinity: pH: 7.82
- Ca/mg ratio: total hardness: 145 CaCO3/L
- Culture medium different from test medium: No
- Intervals of water quality measurement:NS

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light, 8 hours dark
- Light intensity: 1000 - 1500 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality of fish was recorded at 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS

- Range finding study
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: 100 mortality at 10 and 100 mg/L after 24 h; No mortality at 1 mg/L

Reference substance (positive control):

yes

Remarks:

K2Cr2O7 was used as the positive control in the same conditions as the main experiment.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.14 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

During the test period, all fish in the control and 1.0 mg/L treatments were alive and appeared normal. At 2.0 mg/L effects were observed (fish lying on their side). At 4.0 mg/L (measured 3.94 mg/L) all fish were dead after 96 hours. The 96 h LC50 for the rare minnow (Gobiocyprus rarus) was established as 2.14 mg/L (95% confidence interval 1.73 - 2.66 mg/L). The highest concentration causing no mortality (96h LC0) was 1.0 mg/L (measured 0.805 mg/L). The results can be seen in the Any other information on results incl. tables section.


- Other abnormalities: None
- Observations on body length and weight: Not applicable
- Other biological observations: See above
- Mortality of control: None
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
- Effect concentrations exceeding solubility of substance in test medium:

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Mortality: As expected
- LC50: 291 mg/L (24 h) (within the acceptable range of 200-400 mg/L)

Reported statistics and error estimates:

Trimmed Spearman-Karber Method 9Version 1.5, USEPA) was used to calculate the LC50 and 95% confidence limits

Sublethal observations / clinical signs:

Table 1. Mortality during definitive test
Nominal Concentration (mg/L)	Measured Concentration (mg/L)*	Initial Number of Fish	Number of Dead Fish
			3 h	6 h	24 h	48 h	72 h	96 h
0 (Blank control)	ND	10	0	0	0	0	0	0
1	0.805	10	0	0	0	0	0	0
2	1.84	10	0	0	3	3	3	4
4	3.94	10	5	7	10	10	10	10
6	5.82	10	6	10	10	10	10	10
8	7.80	10	10	10	10	10	10	10
10	9.86	10	10	10	10	10	10	10
LC50 (mg/L, based on measured concentrations)					2.30	2.30	2.30	2.14
95% confidence limit (mg/L, based on measured concentrations)					1.88 – 2.81	1.88 – 2.81	1.88 – 2.81	1.73 – 2.66
* - average of measured concentrations at 0, 24, 48, 72, 96 hours

Validity criteria fulfilled:

yes

Conclusions:

Under valid test conditions, the 96 hour LC50 of the test substance to Rare minnow (Gobiocyprus rarus) was 2.14 mg/L (95% confidence limit of 1.73 mg/L-2.66 mg/L) (based on measured concentrations). Maximum tested concentration causing no mortality (96 hr LC0) was 0.805 mg/L (nominal concentration 1 mg/L). Minimum concentration causing 100 % mortality was 3.94 mg/L (nominal concentration 4 mg/L). All fish in the control and 1 mg/L treatments remained alive and appeared normal during the test.

Executive summary:

The effects of the test substance on the fish Rare minnow (Gobiocypris rarus) were assessed according to the OECD guideline 203 "Fish, Acute toxicity Test." (1992). 

An initial range finding test was conducted during which five fish were exposed to each 1, 10 and 100 mg/L of the test substance as well as a control. 100% mortality was observed at 10 and 100 mg/L concentrtaions while no mortality was observed at 1 mg/L concnetration. From the results of this experiment, the following nominal concentrations for the definite test were selected: 1, 2, 4, 6, 8, and 10 mg/L. 

A positive control test using the substance K₂Cr₂O₇ was carried out and the resulting 24 hour LC₅₀ was 291 mg/L, which met the acceptability criteria. 

In the definite test, 10 fish were exposed to each treatment and control, there were no replicates. Samples from control and test treatments were analysed using HPLC-MS and the following mean measured concentrations were obtained: 0.805, 1,84, 3.94, 5.82, 7.80 and 9.86 mg/L, respectively. The deviations were within 20% of the nominal.  The pH values were between 7.67 and 7.92, the dissolved oxygen values varied between 73 and 92% of air saturation at test temperature, which varied betwen 23 and 23.2 degrees Celsius. The observed effects were fish mortality (fish lying on their side and backs). 

The results showed that the 96 hour LC₅₀ of the test substance to Rare minnow (Gobiocyprus rarus) was 2.14 mg/L (95% confidence limit of 1.73 mg/L - 2.66 mg/L) (based on measured concentrations). Maximum tested concentration causing no mortality (96 hr LC₀) was 0.805 mg/L (nominal concentration 1 mg/L). Minimum concentration causing 100% mortality was 3.94 mg/L (nominal concentration 4 mg/L). All fish in the control and 1 mg/L treatments remained alive and appeared normal during the test.

Description of key information

96 h LC50: 2.14 mg/L (Gobiocypris rarus); OECD 203 (1992); Guixiang, J. (2015)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

2.14 mg/L

Additional information

In the key study, the effects of the test substance on the fish Rare minnow (Gobiocypris rarus) were assessed according to the OECD guideline 203 "Fish, Acute toxicity Test." (1992). 

An initial range finding test was conducted during which five fish were exposed to each 1, 10 and 100 mg/L of the test substance as well as a control. 100% mortality was observed at 10 and 100 mg/L concentrtaions while no mortality was observed at 1 mg/L concnetration. From the results of this experiment, the following nominal concentrations for the definite test were selected: 1, 2, 4, 6, 8, and 10 mg/L. 

A positive control test using the substance K₂Cr₂O₇ was carried out and the resulting 24 hour LC₅₀ was 291 mg/L, which met the acceptability criteria. 

In the definite test, 10 fish were exposed to each treatment and control, there were no replicates. Samples from control and test treatments were analysed using HPLC-MS and the following mean measured concentrations were obtained: 0.805, 1,84, 3.94, 5.82, 7.80 and 9.86 mg/L, respectively. The deviations were within 20% of the nominal.  The pH values were between 7.67 and 7.92, the dissolved oxygen values varied between 73 and 92% of air saturation at test temperature, which varied betwen 23 and 23.2 degrees Celsius. The observed effects were fish mortality (fish lying on their side and backs). 

The results showed that the 96 hour LC₅₀ of the test substance to Rare minnow (Gobiocyprus rarus) was 2.14 mg/L (95% confidence limit of 1.73 mg/L - 2.66 mg/L) (based on measured concentrations). Maximum tested concentration causing no mortality (96 hr LC₀) was 0.805 mg/L (nominal concentration 1 mg/L). Minimum concentration causing 100% mortality was 3.94 mg/L (nominal concentration 4 mg/L). All fish in the control and 1 mg/L treatments remained alive and appeared normal during the test.