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EC number: 482-110-2 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
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- Density
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- Endpoint summary
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- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
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Toxicity to aquatic algae and cyanobacteria
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- January 7, 2004 to April 20, 2004
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Version / remarks:
- 1984
- Deviations:
- no
- GLP compliance:
- yes
- Remarks:
- OECD GLP
- Analytical monitoring:
- yes
- Details on sampling:
- Analytical determination of test substance concentration (active ingredient) was performed with samples collected from stock solutions prepared at each concentration prior to their distribution to test vessels at the start of the toxicity test and from each replicate test vessel after 72 hours. Samples collected from replicate test vessels at the end of the test were pooled.
- Vehicle:
- no
- Details on test solutions:
- The dilution water used for acclimation of test organisms and for all toxicity testing was sterile freshwater AAP medium at a pH of 7.5 ± 0.1 (the pH was adjusted with 0.1N NaOH). A sample of the dilution water used for this test contained 1.3 mg/L total organic carbon.
A stock solution with a nominal concentration of 800 mg a.i./L was prepared (with correction for purity) by bringing 1.2575 g of test substance to a total volume of 1,000 mL with algal medium in a glass volumetric flask. This stock solution was used to prepare 13, 25, 50, 100, 200, 400, and 800 mg a.i./L test solutions by transferring the appropriate amount of stock solution into algal medium (the control was 400 mL of algal medium). The pH of the solutions was adjusted to 7.5 ± 0.1. A stability blank sample was set up at 800 mg a.i./L and placed among the test vessels. No algae were inoculated into the stability blank. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- Algae used for the test (Selenastrum capricornutum, UTEX 1648) were from a culture originally procured from the Culture Collection of Algae at the University of Texas at Austin. The culture was transferred to sterile enriched media identical to media used for this test and maintained at test conditions for at least 14 days before the definitive test. During the 14-day acclimation period prior to the start of the test, the temperature ranged from 23.6 to 24.7 °C and the light intensity was 50 to 51 microEin/m²sec (24 hours light/0 hours dark photoperiod).
During the acclimation period, the culture was actively growing in at least two subcultures prior to the start of the toxicity test. The subsample of algae used to inoculate media at the start of the definitive test came from a four day old culture. Identification of the culture organisms, which are also referred to as Raphidocelis subcapitata, was verified using an appropriate taxonomic key. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Test temperature:
- 23.4 to 23.7 °C
- pH:
- 7.5 to 7.6 (initial); 7.2 to 9.3 (termination)
- Nominal and measured concentrations:
- Nominal: 0 mg a.i./L (control), 13, 25, 50, 100, 200, 400 and 800 mg/L
Mean Measured: ND (none detected, control), 12, 23, 47, 95, 190, 380 and 790 mg a.i./L - Details on test conditions:
- A range-finding test was conducted with a control and five concentrations of the test substance: 5.0, 10, 50, 100, and 500 mg a.i./L. At the conclusion of the test, the number of cells/mL in the test vessels equalled the following percent of the number of cells/mL in the control vessels: 5.0 and 10 mg a.i./L = greater than 95%, 50 mg a.i./L = 35%, 100 mg a.i./L = 14%, and 500 mg a.i./L = 9%. Insoluble material was not observed. A definitive test was conducted with a control and five concentrations of the test substance: 13, 25, 50, 100, and 200 mg a.i./L. At the conclusion of the test, the number of cells/mL in the test vessels equalled the following percent of the number of cells/mL in the control vessels: 13 mg a.i./L = 91%, 25 mg a.i./L = 70%, 50 mg a.i./L = 52%, 100 mg a.i./L = 35%, and 200 mg a.i./L = 14%. Insoluble material was not observed. The test was repeated with higher concentrations to allow EC50 calculations using the average specific growth rate.
The final definitive toxicity test was conducted at 24 ± 2°C with seven concentrations of test substance and a control. Water quality measurements were made and each solution was subdivided into clear glass 250 mL Erlenmeyer flasks (three replicates of the control and each test concentration). Flasks were inoculated with approximately 10,000 algal cells/mL and capped with clear glass inverted beakers. Test vessels were randomly arranged on a rotary shaker adjusted to approximately 100 rpm in an incubator during the test (a random numbers table was used to select the location for each vessel) and repositioned by five places each day. A 24 hour light and 0 hour dark photoperiod was automatically maintained with cool white fluorescent lights that provided a light intensity of approximately 370 to 390 footcandles (also measured as approximately 51 to 53 microEin/m²sec).
The number of algal cells/mL in each test vessel and the occurrence of relative size differences, unusual cell shapes, colors, flocculations, adherence of cells to test containers, or aggregation of cells was determined visually by means of direct microscopic examination with a hemocytometer at 24, 48, and 72 hours. Temperature of the incubator was measured and recorded daily and the temperature in a representative vessel of water incubated with the test vessels was continuously recorded with noted interruption(s). The pH of test solutions was measured and recorded in each test solution prior to distribution to the test vessels at the start of the test and in each test vessel at the end of the test.
At the conclusion of the toxicity test, a determination of whether the toxic effects observed at 95, 190, 380, and 790 mg a.i./L were algistatic or algicidal was made by transferring 0.50 mL of test solution from each flask to 100 mL of dilution water without the test substance and incubating these flasks under test conditions for 96 to 120 hours. At the conclusion of this incubation period, the number of cells/mL was determined in each flask. - Duration:
- 24 h
- Dose descriptor:
- EC50
- Effect conc.:
- 620 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 500-780 mg/L
- Duration:
- 48 h
- Dose descriptor:
- EC50
- Effect conc.:
- 280 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 220-370 mg/L
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 180 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 140-250 mg/L
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 47 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- act. ingr.
- Basis for effect:
- growth rate
- Details on results:
- Insoluble material was not observed during the definitive toxicity test. The mean measured concentrations were 92 to 99% of nominal concentrations, indicating that the test substance was stable under test conditions. All results were based on mean measured test concentrations. The algal population grew well, resulting in an average of 768,000 cells/mL in the control after 72 hours. No effects (relative size differences, unusual cell shapes, colors, flocculations, adherence of cells to test containers, or aggregation of cells) were noted during the test. Water quality throughout the test was within acceptable limits. The incubator temperature ranged from 23.4 to 23.7°C.
At the conclusion of the final definitive toxicity test, a 0.5 mL aliquot of test media from the 95, 190, 380, and 790 mg a.i./L concentrations was combined with 100 mL of fresh media and incubated under test conditions. During a 96-hour incubation period, the algal population increased from approximately 3,500 cells/mL to 726,000 cells/mL at 95 mg a.i./L and from approximately 890 cells/mL to 206,000 cells/mL at 190 mg a.i./L. During a 120-hour incubation period, the algal population grew from approximately 620 cells/mL to 214,000 cells/mL at 380 mg a.i./L and from approximately 470 cells/mL to 154,000 cells/mL at 790 mg a.i./L, indicating that the toxic effects were algistatic rather than algicidal. - Reported statistics and error estimates:
- The average specific growth rate was calculated as the natural log of the number of cells/mL at time t1 minus the natural log of the number of cells/mL at time t0 divided by the exposure period. The 24, 48, and 72 hour EC50 values were calculated using a weighted least squares non-linear regression technique, when possible. The slope of the concentration-response curve is not calculated by this method. The no observed effect concentration (NOEC) was determined using a one-way analysis of variance (ANOVA) and Bonferroni’s test. The NOECs were determined using the mean measured concentrations of the test substance, and the number of cells/mL and average specific growth rate. Treatment data were compared to control data for the determination of the NOECs.
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study exposure of algae to to the test material for 72 hours resulted in an EC50 of 58 mg/L (95 % confidence interval = 40 to 85 mg/L) when determined using the number of cells/mL and 180 mg/L (95 % confidence interval = 140 to 250 mg/L) when determined using the average specific growth rate. When treatment data are compared to the control, the 72 hour NOEC is 12 mg/L test material when determined using the number of cells/mL and 47 mg/L when determined using the average specific growth rate.
- Executive summary:
The acute toxicity of the test material to freshwater algae, Selenastrum capricornutum, was investigated according to the standardised guidelines OECD 201, under GLP conditions.
The test was performed under static conditions with seven concentrations of test material and a dilution water control at 24 ± 2 °C. Nominal concentrations of the test material were 0 mg/L (control), 13, 25, 50, 100, 200, 400, and 800 mg/L. Mean measured concentrations were ND (none detected; control), 12, 23, 47, 95, 190, 380, and 790 mg/L. The mean measured concentrations were 92 to 99 % of nominal concentrations, indicating that the test material was stable under test conditions. Insoluble material was not observed at any tested concentration during the test.
The dilution water was sterile freshwater AAP medium adjusted to a pH of 7.5 ± 0.1. The test was performed in clear glass 250 mL flasks that contained 100 mL of test solution. Algae used in the test were acclimated to test conditions for more than 14 days. Algae were distributed among three replicates of the control and each test concentration at the rate of approximately 10,000 cells/mL.
Under the conditions of this study exposure of algae to to the test material for 72 hours resulted in an EC50 of 58 mg/L (95 % confidence interval = 40 to 85 mg/L) when determined using the number of cells/mL and 180 mg/L (95 % confidence interval = 140 to 250 mg/L) when determined using the average specific growth rate. When treatment data are compared to the control, the 72 hour NOEC is 12 mg/L test material when determined using the number of cells/mL and 47 mg/L when determined using the average specific growth rate.
Reference
Measured concentrations of the test substance during the toxicity test with the freshwater alga, Selenastrum capricornutum.
Nominal Concentration of the Test Substance (mg a.i./L) |
Measured Concentrations of the Test Substance |
|||
0 Hour |
72 Hour |
Mean |
% Recovery |
|
Test Media Samples |
||||
Control |
ND |
ND |
ND |
-- |
13 |
12 |
11 |
12 |
92 |
25 |
23 |
23 |
23 |
92 |
50 |
47 |
47 |
47 |
94 |
100 |
95 |
94 |
95 |
95 |
200 |
190 |
190 |
190 |
95 |
400 |
380 |
380 |
380 |
95 |
800 |
800 |
770 |
790 |
99 |
Stability Blank |
||||
800 |
800 |
820 |
810 |
101 |
Matrix Spike Blank |
||||
100 |
-- |
100 |
-- |
-- |
-- |
100 |
100 |
100 |
|
Laboratory Control Spike Sample |
||||
100 |
99 |
99 |
99 |
99 |
Blank |
||||
0 |
ND |
ND |
ND |
-- |
ND = none detected at or above the limit of quantitation |
Cell growth data from the toxicity test with the test substance and the freshwater alga, Selenastrum capricornutum.
Mean Measured Conc. of the Test Substance |
R |
Number of Cells per Milliliter |
|||
Hour of Exposure |
|||||
0 |
24 |
48 |
72 |
||
Control |
1 |
10,000 |
46,000 |
200,000 |
802,000 |
2 |
10,000 |
49,000 |
246,000 |
740,000 |
|
3 |
10,000 |
48,000 |
216,000 |
762,000 |
|
Mean |
10,000 |
48,000 |
221,000 |
768,000 |
|
12 |
1 |
10,000 |
56,000 |
236,000 |
820,000 |
2 |
10,000 |
62,000 |
206,000 |
766,000 |
|
3 |
10,000 |
60,000 |
210,000 |
810,000 |
|
Mean |
10,000 |
59,000 |
217,000 |
799,000 |
|
% control |
100 |
123 |
98 |
104 |
|
23 |
1 |
10,000 |
48,000 |
144,000 |
630,000 |
2 |
10,000 |
58,000 |
168,000 |
672,000 |
|
3 |
10,000 |
66,000 |
188,000 |
684,000 |
|
Mean |
10,000 |
57,000 |
167,000 |
662,000 |
|
% control |
100 |
119 |
76 |
86 |
|
47 |
1 |
10,000 |
39,000 |
166,000 |
502,000 |
2 |
10,000 |
50,000 |
146,000 |
676,000 |
|
3 |
10,000 |
47,000 |
150,000 |
612,000 |
|
Mean |
10,000 |
45,000 |
154,000 |
597,000 |
|
% control |
100 |
94 |
70 |
78 |
|
95 |
1 |
10,000 |
45,000 |
90,000 |
192,000 |
2 |
10,000 |
50,000 |
120,000 |
260,000 |
|
3 |
10,000 |
48,000 |
118,000 |
238,000 |
|
Mean |
10,000 |
48,000 |
109,000 |
230,000 |
|
% control |
100 |
100 |
49 |
30 |
|
190 |
1 |
10,000 |
50,000 |
54,000 |
54,000 |
2 |
10,000 |
33,000 |
62,000 |
58,000 |
|
3 |
10,000 |
46,000 |
60,000 |
66,000 |
|
Mean |
10,000 |
43,000 |
59,000 |
59,000 |
|
% control |
100 |
90 |
27 |
8 |
|
380 |
1 |
10,000 |
21,000 |
39,000 |
40,000 |
2 |
10,000 |
42,000 |
32,000 |
34,000 |
|
3 |
10,000 |
33,000 |
38,000 |
48,000 |
|
Mean |
10,000 |
32,000 |
36,000 |
41,000 |
|
% control |
100 |
67 |
16 |
5 |
|
790 |
1 |
10,000 |
20,000 |
17,000 |
31,000 |
2 |
10,000 |
21,000 |
34,000 |
27,000 |
|
3 |
10,000 |
18,000 |
28,000 |
35,000 |
|
Mean |
10,000 |
20,000 |
26,000 |
31,000 |
|
% control |
100 |
42 |
12 |
4 |
R= replicate
Average specific growth rate from the toxicity test with the test substance and the freshwater alga, Selenastrum capricornutum.
Mean Measured Conc. of the Test Substance (mg a.i./L) |
|
Average Specific Growth Rate |
||
Hours of Exposure |
||||
24 |
48 |
72 |
||
Control |
Mean |
0.065 |
0.064 |
0.060 |
12 |
Mean |
0.074 |
0.064 |
0.061 |
% control |
114 |
100 |
102 |
|
23 |
Mean |
0.073 |
0.059 |
0.058 |
% control |
112 |
92 |
97 |
|
47 |
Mean |
0.063 |
0.057 |
0.057 |
% control |
97 |
89 |
95 |
|
95 |
Mean |
0.065 |
0.050 |
0.044 |
% control |
100 |
78 |
73 |
|
190 |
Mean |
0.061 |
0.037 |
0.025 |
% control |
94 |
58 |
42 |
|
380 |
Mean |
0.048 |
0.027 |
0.020 |
% control |
74 |
42 |
33 |
|
790 |
Mean |
0.029 |
0.020 |
0.016 |
% control |
45 |
31 |
27 |
Description of key information
Ward, Wyskiel, Boeri (2003)
Under the conditions of this study exposure of algae to to the test material for 72 hours resulted in an EC50 of 58 mg/L (95 % confidence interval = 40 to 85 mg/L) when determined using the number of cells/mL and 180 mg/L (95 % confidence interval = 140 to 250 mg/L) when determined using the average specific growth rate. When treatment data are compared to the control, the 72 hour NOEC is 12 mg/L test material when determined using the number of cells/mL and 47 mg/L when determined using the average specific growth rate.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 180 mg/L
- EC10 or NOEC for freshwater algae:
- 47 mg/L
Additional information
Ward, Wyskiel, Boeri (2003)
The acute toxicity of the test material to freshwater algae, Selenastrum capricornutum, was investigated according to the standardised guidelines OECD 201, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).
The test was performed under static conditions with seven concentrations of test material and a dilution water control at 24 ± 2 °C. Nominal concentrations of the test material were 0 mg/L (control), 13, 25, 50, 100, 200, 400, and 800 mg/L. Mean measured concentrations were ND (none detected; control), 12, 23, 47, 95, 190, 380, and 790 mg/L. The mean measured concentrations were 92 to 99 % of nominal concentrations, indicating that the test material was stable under test conditions. Insoluble material was not observed at any tested concentration during the test.
The dilution water was sterile freshwater AAP medium adjusted to a pH of 7.5 ± 0.1. The test was performed in clear glass 250 mL flasks that contained 100 mL of test solution. Algae used in the test were acclimated to test conditions for more than 14 days. Algae were distributed among three replicates of the control and each test concentration at the rate of approximately 10,000 cells/mL.
Under the conditions of this study exposure of algae to to the test material for 72 hours resulted in an EC50 of 58 mg/L (95 % confidence interval = 40 to 85 mg/L) when determined using the number of cells/mL and 180 mg/L (95 % confidence interval = 140 to 250 mg/L) when determined using the average specific growth rate. When treatment data are compared to the control, the 72 hour NOEC is 12 mg/L test material when determined using the number of cells/mL and 47 mg/L when determined using the average specific growth rate.
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