Amines, N-C10-16-alkyltrimethylenedi-, reaction products with chloroacetic acid

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish, juvenile growth test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-06-28 to 2007-07-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 215 (Fish, Juvenile Growth Test)

Version / remarks:

adopted 21 January 2000

Deviations:

yes

Remarks:

Two specimen, exceeded the threshhold value of 25% by 1% and 2 %, respectively. However, fish length was in the recommended range of ±10%. Analytical verification was only performed at the two highest test concentrations.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 50 mL samples of each test concentration (only concentrations 37.5 and 75.0 a.i. were analyzed)
- Sample storage conditions before analysis: stored frozen at -18°C until analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock-solution was prepared by mixing the test substance in the dilution water. The stock solution was continually dispensed and diluted with dilution water using the respective equipment to deliver a series of five concentrations at the same time to the corresponding test vessels.
- Controls: Dilution water only

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: Eye point stage eggs were obtained from the NRW Landesanstalt für Fischerei (governmental fisheries agency), Albaum, Germany on March 22, 2007 and further bred in the test facility.
- Length and weight at study initiation: 5 cm ± 1 cm; 3 g ± 1 g
- Method of breeding: Rainbow trout fingerlings were bred at 12.0 °C under flowthrough conditions in the laboratory of the Fh-IME. During the week the trouts were fed daily with commercial food. Larvae were fed daily ad libitum from the day of the first swim-up with pulverized trout food (Trouvit Alleinfuttermittel, Milkivit, D86664 Burgheim). From day 17 on, crushed trout food was given.

Only fish without visible abnormalities were used in the study.

FEEDING DURING TEST
- Food type: Trout food (Trouvit Alleinfuttermittel, Milkivit, D- 86664 Burgheim)
- Amount: All fish were weighed before test start. A feeding rate of 4 % of body weight was adjusted, divided into two equal portions per day.
The fish were weighed again on day 14 of the test to recalculate and adjust the feeding rate, and on day 28 to obtain the results of growth.
- Frequency: Twice per day

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

28 d

Test temperature:

13.3–14.9 °C

pH:

7.5–8.1

Dissolved oxygen:

65–98 %

Nominal and measured concentrations:

Nominal: 23.5, 42.0, 93.5, 188 and 375 μg/L (corresponding to nominal concentrations of 4.7, 9.4, 18.8, 37.5 and 75.0 μg active substance/L)
Due to a complete lack of adverse effects on juvenile growth, only the two highest test item concentrations (37.5 and 75 μg/L, nominal) were verified analytically.
The arithmetic means of the measured concentrations of the four lead compounds were extrapolated to the total content of active substance.
The recovery rate at 37.5 μg a.s./L (nominal) was 36.3 %, thus resulting in an actual test item concentration of 13.6 µg a.s./L.
At a nominal test item concentration of 75 μg a.s./L, 69.7 % of the lead compounds were recovered, corresponding to 52.3 µg a.s./L.

Details on test conditions:

TEST SYSTEM
- Type: open
- Material, fill volume: glass aquaria, 25 L
- Aeration: yes
- Renewal rate of test solution: Flow rate of about 2.5 L/h (daily turnover of about 5 volumes)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: < 2.0 g fish wet weight/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Cu-free tap water
- Alkalinity: 0.7–1.1 mmol/L
- hardness: 0.7–1.0 mmol total hardness
- pH: 8.3
- Oxygen content: 80 % oxygen saturation
- Conductivity: 161.3–183.7
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light/8 h dark
- Light intensity: did not exceed 1000 lux

EFFECT PARAMETERS MEASURED:
Behaviour and mortality: Daily
Weight and length: Test start and after 14 days and 28 days
Analytical samples: Test start and days 7, 14, 21, and 28

Reference substance (positive control):

no

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 52.3 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

other: Mortality, wet weight, body length, pseudo-specific growth rate

Details on results:

No abnormal condition or behaviour was observed at any test concentration up to and including 52.3 µg/L (highest concentration tested, mean measured value).
During the test no test item related mortality occurred. One fish was lost due to handling. At any concentration up to and including 52.3 µg a.s./L (measured), neither any significant effect nor any trend was observed.
The fish weights increased by approx. 100 % every 14 d, thus the quality criterion for the controls (at least 50% in 28 d) was met even by all treatments.
Regarding the pseudo-specific growth rate, no test item related effect was observed. The NOEC was established at ≥ 52.3 µg a.s./L (measured).
Summary of the effect data is given in Table A7.4.3.2- 9

Reported statistics and error estimates:

Since no effect on mortality, weight and pseudo-specific growth rate occurred, no statistical evaluation was performed.

Table A7.4.3.2-5: Measured concentrations and mean of the four “Lead components” of the active substance in the "flow-through Oncorhynchus mykiss, juvenile growth test" test media (75.0 µg a.s./L, nominal).

Lead compound	Nominal concentration [µg/L]	Measured [µg/L] (% nom.)
		Test start	7 d	14 d	21 d	28 d	Mean
C-12 PDA	26.71	29.95 (112.1)	15.76 (59.0)	16.29 (61.0)	9.34 (35.0)	3. 74 (14.0)	15.01 (56.21)
N-C12 Gly	7.8	9.58 (122.8)	8.03 (103.0)	15.73 (201.7)	16.75 (214.7)	6.51 (83.5)	11.32 (145.12)
N'-C12 Gly	11.70	6.32 (54.0)	4.53 (38.7)	7.94 (67.9)	8.77 (75.0)	3.36 (28.7)	6.19 (52.86)
N'-C12 diGly	3.23	1.65 (51.1)	1.40 (43.3)	2.63 (81.5)	3.14 (97.1)	0.98 (30.4)	1.96 (60.71)
Sum of lead components	49.44	47.49 (96.0)	29.73 (60.1)	42.60 (86.2)	37.99 (76.8)	14.59 (29.5)	34.48 (69.74)
Extrapolated to total a.s.	75.0						52.3 (69.7)

 

Table A7.4.3.2-6: Temperature [°C] in the test tanks throughout the test. a.s. = active substance; n. = nominal; SD = standard deviation; m.m. = mean measured; Concentrations given as nominal and mean measured concentrations, where appropriate.

	a.s. concentration [µg/L]
	Control	4.69 n.	9.38 n.	18.75 n.	37.50 n.	75.00 n. (52.3 m.m.)
Min	13.5	13.4	13.5	13.3	13.4	13.4
Max	14.9	14.6	14.5	14.3	14.3	14.6
Mean	14.3	14.1	14.0	13.9	13.9	14.2
SD	0.3	0.3	0.3	0.2	0.3	0.3

 

Table A7.4.3.2-7: Oxygen saturation of the test media (%).Values of the test tanks throughout the test (28 days). a.s. = active substance; n. = nominal; SD = standard deviation; m.m. = mean measured; Concentrations given as nominal and mean measured concentrations, where appropriate.

	a.s. concentration [µg/L]
	Control	4.69 n.	9.38 n.	18.75 n.	37.50 n.	75.00 n. (52.3 m.m.)
Min	70	70	65	66	66	70
Max	98	91	95	90	95	98
Mean	81	81	81	80	81	80
SD	7	6	8	6	5	7

 

Table A7.4.3.2-8:pH of the test media in the test tanks throughout the test (28 days). a.s. = active substance; n. = nominal; SD = standard deviation; m.m. = mean measured; Concentrations given as nominal and mean measured concentrations, where appropriate.

	a.s. concentration [µg/L]
	Control	4.69 n.	9.38 n.	18.75 n.	37.50 n.	75.00 n. (52.3 m.m.)
Min	7.57	7.54	7.52	7.51	7.58	7.57
Max	8.08	8.04	8.05	8.08	8.09	8.09
Mean	7.8	7.8	7.8	7.7	7.7	7.7
SD	0.2	0.1	0.1	0.1	0.1	0.1

 

Table A7.4.3.2-9:Effect data: Fish weights (g) and length (cm) at test start, day 14 and at test end (28 days). (n per vessel = 10). a.s. = active substance; SD = standard deviation; n. = nominal; m.m. = mean measured; concentrations given as nominal and mean measured concentrations; mean individual pseudo-specific r₃(tank-average specific r₂) growth rate from day 0 to 28. (n per vessel = 10); no significant deviation occurred, compared to the control.

		a.s. concentration [µg/L]
		Control	4.69 n.	9.38 n.	18.75 n.	37.50 n.	75.00 n. (52.3 m.m.)
Fish weights	Mean 28 d	8.33 g (SD 1.64)	8.69 g (SD 1.13)	8.5 g (SD 2.11)	8.44 g (SD 1.32)	8.7 g (SD 1.89)	8.49 g (SD 2.09)
	Mean gain* 0–28 d	5.75 g	5.95 g	5.7 g	5.75 g	5.83 g	5.78 g
Fish length	Mean 28 d	8.9 cm (SD 0.5)	8.8 cm (SD 0.5)	8.8 cm (SD 0.6)	8.7 cm (SD 0.5)	8.9 cm (SD 0.6)	8.7 cm (SD 0.7)
	Mean gain* 0–28 d	2.5	2.5	2.4	2.4	2.5	2.4
Pseudo-specific growth rate	Mean (=r2)	4.2 (SD 0.7)	4.3 (SD 0.5)	4.2 (SD 0.9)	4.2 (SD 0.6)	4.3 (SD 0.8)	4.2 (SD 0.8)
No fish showed any clinical sign of intoxication * pseudo-specific weight/length gain as individual weight after 28 d / mean weight at test start

 

 

Validity criteria fulfilled:

yes

Conclusions:

The NOEC of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid in juvenile rainbow trout (Oncorhynchus mykiss) was determined to be ≥ 52.3 μg a.i./L; no effects were observed up to and including the highest tested concentration.

Executive summary:

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid (20% a.i., as manufactured) on growth and survival of juvenile rainbow trout (Oncorhynchus mykiss) was tested according to OECD guideline 215 (2000) at nominal concentrations of 4.7, 9.4, 18.8, 37.5 and 75.0 μg active substance/L under flow-through conditions for 28 days.

At mean measured concentrations up to and including 52.3μg a.s./L, neither any significant effect nor any trend was observed. The fish grew well (203–223% of start weight) during the test and fulfilled the control validity criteria of the OECD Guideline 215 at all treatment levels. As no effect was observed, the NOEC of the test item related to mean measured concentration was determined to be ≥ 52.3 μg a.s./L, representing ≥ 261.5 μg test item per litre.

Description of key information

NOEC (28 d) = ≥ 52.3 µg a.i./L (mean measured, Oncorhynchus mykiss, OECD Guideline 215)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

52.3 µg/L

Additional information

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid (20% a.i., as manufactured) on growth and survival of juvenile rainbow trout (Oncorhynchus mykiss) was tested according to OECD guideline 215 (2000) at nominal concentrations of 4.7, 9.4, 18.8, 37.5 and 75.0 μg a.i./L under flow-through conditions for 28 days.

At mean measured concentrations up to and including 52.3 μg a.i./L, neither any significant effect nor any trend was observed. The fish grew well (203–223% of start weight) during the test and fulfilled the control validity criteria of the OECD Guideline 215 at all treatment levels. As no effect was observed, the NOEC of the test item related to mean measured concentration was determined to be ≥ 52.3 μg a.i./L, representing ≥ 261.5 μg test item per litre.