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EC number: 405-520-5 | CAS number: 95235-30-6 D-8; DD-8
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- 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
Long-term toxicity to aquatic invertebrates
Administrative data
Link to relevant study record(s)
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- from 1993-03-01 to 1993-12-02
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: OECD 202: Daphnia sp.; Acute Immobilisation Test and Reproduction Test, Part 2
- Version / remarks:
- 04 April 1984
- Deviations:
- no
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 211 (Daphnia magna Reproduction Test)
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- Unfiltered samples (20 mL) of aqueous test media were diluted to equate HPLC mobile phase to within the nominal concentration range of 0.1 to 10 mg/L. The concentrations of D-8 in the diluted samples were then determined by high performance liquid chromatography using a spectrophotometric detector.
- Vehicle:
- no
- Details on test solutions:
- Two aqueous stock preparations of D-8 were prepared at nominal concentrations of 5 and 15 mg/L. Test material was directly added to the dilution water. As the 15 mg/L concentration is near the aqueous solubility limit of D-8 (19.7 mg/L), the stock solutions were shaken overnight to ensure complete dissolution. These stock solutions were used directly at the two highest test levels. The 5 mg/L stock solution was then diluted with water to provide the three lower test levels (0.15 to 1.5 mg/L).
- Test organisms (species):
- Daphnia magna
- Details on test organisms:
- TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: genetically homogeneous
- Source: University of Sheffield; England
- Age of parental stock:
- Feeding during test
- Food type: Algae (Chlorella vulgaris) and baker's yeast
- Amount: algae density: 2 to 8E5 cells per mL; yeast: suspension with a nominal concentration of 0.04 to 0.05 mg/L
- Frequency: five times each week
ACCLIMATION
- Acclimation period: not indicated
- Acclimation conditions: not indicated - Test type:
- semi-static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 21 d
- Hardness:
- 45 to 59 mg/L as CaCO3
- Test temperature:
- 20.0 ± 0.7 °C
- pH:
- 7.1 to 8.2
- Dissolved oxygen:
- 95 - 103 % ASV (ASV = Air Saturation Value)
- Nominal and measured concentrations:
- Nominal: 0.15; 0.5; 1.5; 5 and 15 mg/L
Measured: 0.139; 0.492; 1.45; 5.04 and 14.8 mg/L - Details on test conditions:
- Groups of forty Daphnia were exposed to D-8 at nominal concentrations of 0.15, 0.5, 1,5, 5 and 15 mg/L. Five Daphnia were placed into each of the test vessels. Test media were renewed three times each week. A control group of Daphnia was exposed to dilution water alone. The test was conducted at 20 ± 0.7 °C in treated tap water of hardness 45 to 59 mg/L as CaCO3 at pH 7.1 to 8.2. At all concentrations, test media were clear and colourless on days of preparation. Exposure levels were monitored by HPLC method. The numbers of mobile, immobile and dead parental Daphnia were recorded each day of the test and the numbers of juveniles produced were counted three times each week.
A 48-hour range finding toxicity test showed an EC50 between 10 and 16 mg/L. The limit of water solubility of D-8 is 19.7 mg/L. - Reference substance (positive control):
- no
- Key result
- Duration:
- 21 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.492 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- mean cumulative number of juveniles per surviving adult
- Key result
- Duration:
- 21 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.139 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- total number of juveniles per vessel
- Key result
- Duration:
- 21 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.139 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Duration:
- 21 d
- Dose descriptor:
- LC50
- Effect conc.:
- 2.7 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Remarks on result:
- other: 95 % CI: 1.80 - 3.65 mg/L
- Duration:
- 14 d
- Dose descriptor:
- LC50
- Effect conc.:
- 5.4 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Remarks on result:
- other: approximate values
- Duration:
- 7 d
- Dose descriptor:
- LC50
- Effect conc.:
- 7.4 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Remarks on result:
- other: 95 % CI: 5.91 - 8.89 mg/L
- Duration:
- 96 h
- Dose descriptor:
- LC50
- Effect conc.:
- 10.5 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Remarks on result:
- other: 95 % CI: 9.02 - 12.5 mg/L
- Duration:
- 48 h
- Dose descriptor:
- LC50
- Effect conc.:
- 13.4 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Remarks on result:
- other: approximate values
- Duration:
- 21 d
- Dose descriptor:
- EC50
- Effect conc.:
- 1.9 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- total number of juveniles produced at each meas. concentration
- Duration:
- 21 d
- Dose descriptor:
- EC50
- Effect conc.:
- 2.94 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Remarks:
- mean cumulative number of juveniles per adult
- Details on results:
- Mortality of parental Daphnia:
After 21 days, 17.5 % of the parental control Daphnia had died. In groups exposed to D-8, mortality was concentration-related, ranging from 22.5 % at the lowest exposure level (0.139 mg/L) to 100 % at the highest (14.8 mg/L).
Juvenile production:
On Day 5, gravid Daphnia were observed in all of the control vessels, in all of the test vessels at 0.139, 0.492 and 1.45 mg/L and in two of the vessels at 5.04 mg/L. After eight days, the majority of the Daphnia in the control vessels and in the test vessels up to and including 1.45 mg/L were gravid, and at 5.04 mg/L each vessel contained some gravid Daphnia. No gravid Daphnia were observed at 14.8 mg/L during the test. - Reported statistics and error estimates:
- Median lethal concentrations (LC50's) for parental generation were calculated by appropriate statistical methods (binomial, moving average or probit). Median effect concentrations (EC50's) for reproduction were similarly calculated using the total number of juveniles produced at each concentration, and the cumulative number of juveniles per adult.
The numbers of dead parental Daphnia, the total number of juveniles and the mean number of juveniles produced per adult in the control group were compared with those in the test groups, by Dunnett's multi-comparison test which uses a multiple t-test with a common estimate of experimental error. - Validity criteria fulfilled:
- yes
- Conclusions:
- D-8 was tested for 21-day juvenile production study in Daphnia magna according to OECD Guideline 202. The 21-day median lethal concentration (LC50) of D-8 to parental Daphnia was calculated to be 2.70 mg/L. The 21-day EC50 values for inhibition of reproduction were 1.90 mg/L based on the total number of juveniles produced at each mean measured concentration, and 2.94 mg/L based on the mean cumulative number of juveniles produced per adult. In conclusion, D-8 significantly affected both the survival of the parental generation and their ability to produce viable juveniles at concentrations above 0.139 mg/L. Therefore, NOEC was determined to be 0.139 mg/L.
- Executive summary:
The effect of D-8 on the survival and reproduction of Daphnia magna has been assessed under semi-static exposure conditions over a period of 21 days in accordance with OECD 202, Part 2 (1984). Groups of forty Daphnia were exposed to D-8 at nominal concentrations of 0.15, 0.5, 1.5, 5 and 15 mg/L, selected following a ten-day range finding test. A control group of Daphnia was exposed to dilution water alone. The test was conducted at 20 ± 0.7 °C in treated tap water of hardness 45 to 59 mg/L as CaCO3 at pH 7.1 to 8.2. Exposure levels were monitored by HPLC analysis. The numbers of mobile, immobile and dead parental Daphnia were recorded each day of the test, and the numbers of juveniles produced were counted three times each week.
In groups exposed to D-8, mortality of the parental Daphnia after 21 days ranged from 22.5 % at the lowest level (0.139 mg/L) to 100 % at the highest level (14.8 mg/L). Mortality of the parental control Daphnia after 21 days was 17.5 %.
The 21-day median lethal concentration (LC50) of D-8 to parental Daphnia was calculated to be 2.70 mg/L, with 95 % confidence limits of 1.80 - 3.65 mg/L.
The 21-day EC50 values for inhibition of reproduction were 1.90 mg/L, based on the total number of juveniles produced at each mean measured concentration, and 2.94 mg/L, based on the mean cumulative number of juveniles produced per adult. In conclusion, D-8 significantly affected both the survival of the parental generation and their ability to produce viable juveniles at concentrations above 0.139 mg/L. Therefore, NOEC was determined to be 0.139 mg/L.
- Endpoint:
- long-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 211 (Daphnia magna Reproduction Test)
- Version / remarks:
- 2012
- Deviations:
- no
- GLP compliance:
- no
- Analytical monitoring:
- yes
- Test organisms (species):
- Daphnia magna
- Details on test organisms:
- TEST ORGANISM
- Common name: D. magna
- Feeding during test
- Food type: Chlorella vulgaris (AquaNet, Gyeongnam, Korea)
- Frequency: daily - Test type:
- not specified
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 21 d
- Hardness:
- monitored daily
- Test temperature:
- monitored daily
- pH:
- monitored daily
- Dissolved oxygen:
- monitored daily
- Salinity:
- monitored daily
- Conductivity:
- monitored daily
- Nominal and measured concentrations:
- 0, 0.06, 0.12, 0.25, 0.5, 1 mg/L (nominal)
- Details on test conditions:
- TEST SYSTEM
- Renewal rate of test solution: at least three times a week
- No. of organisms per vessel: 10
OTHER TEST CONDITIONS
- Photoperiod: 16:8 h light:dark
EFFECT PARAMETERS MEASURED: survival of parent animal, first day of neonate production, number of neonates per parent animal, number of neonates per brood, and growth
- Reference substance (positive control):
- no
- Key result
- Duration:
- 21 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mortality
- Key result
- Duration:
- 21 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- reproduction
- Duration:
- 21 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: body length
- Validity criteria fulfilled:
- not specified
- Conclusions:
- The NOEC value for reproduction of D-8 in this 21-d chronic toxicity test was 1 mg/L, the highest concentration tested.
- Executive summary:
In this study, long-term toxicity tests of D-8 were performed using Daphnia magna. The Daphnia magna (21 d) chronic toxicity tests were conducted as outlined in Organization for Economic Cooperation and Development (OECD) test guidelines 211 (OECD, 2012). For D. magna chronic toxicity tests, 10 neonates per group were exposed to D-8 (0, 0.06, 0.12, 0.25, 0.5, 1 mg/L). The endpoints were survival of parent animal, first day of neonate production, number of neonates per parent animal, number of neonates per brood, and growth. InD. magnaexposed to D-8 for 21 d, no adverse effects of survival and reproduction were observed up to the highest test concentration of 1 mg/L. However, the body length was significantly reduced when exposed to 1 mg/L of D-8. The NOEC value for reproduction of D-8 was 1 mg/L. D-8 was also tested for 7 days in Moina macrocopa, an indigenous species in Asia, in concentrations of 0, 0.01, 0.03, 0.11, 0.33, 1 mg/L. The endpoints were survival of parent animal, first day of neonate production, number of neonates per parent animal, number of neonates per brood. The first day of reproduction was significantly increased at 1 mg/L D-8. The number of young per female, one of the most sensitive endpoints related to the reproduction, was significantly decreased with treatment of 0.3 mg/L of D-8.The NOEC value for reproduction of D-8 was 0.1 mg/L.
Referenceopen allclose all
Table 1. Analytical determination of D-8
Sampling time, days |
Nominal D-8 concentrations, mg/L |
||||
0.15 |
0.5 |
1.5 |
5.0 |
15.0 |
|
Untreated samples |
|||||
5 (new) |
0.154, 0.154 |
0.531, 0.539 |
1.68, 1.56 |
5.20, 5.46 |
14.8, 14.8 |
8 (old) |
0.143, 0.146 |
0.498, 0.499 |
1.52, 1.53 |
5.12, 5.15 |
14.9, 14.9 |
8 (new) |
0.144, 0.140 |
0.510, 0.500 |
1.49, 1.52 |
5.00, 5.03 |
14.7, 14.8 |
11 (old) |
0.133, 0.129 |
0.474, 0.448 |
1.40, 1.38 |
5.06, 4.90 |
14.6, 14.6 |
15 (new) |
0.133, 0.150 |
0.491, 0.494 |
1.34, 1.35 |
5.03, 5.01 |
- |
18 (old) |
0.123, 0.124 |
0.456, 0.460 |
1.28, 1.29 |
4.71, 4.76 |
- |
Mean value |
0.139 |
0.492 |
1.45 |
5.04 |
14.8 |
Centrifuged samples |
|||||
8 (new) |
0.137 |
0.484 |
1.46 |
4.96 |
14.7 |
11 (old) |
0.130 |
0.464 |
1.39 |
4.78 |
14.3 |
Control samples: no D-8 was detected in every measurement of the control samples
(): Mean measured concentrations expressed
as percentage of the nominal levels.
-: No analysis was required at the level because all of the parental
animals died.
Table 2. Cumulative number of dead parental Daphnia
Observation time (days) |
Measured D-8 concentration (mg/L) |
|||||
0 |
0.139 |
0.492 |
1.45 |
5.04 |
14.8 |
|
1 |
0 |
0 |
0 |
0 |
0 |
19 |
2 |
0 |
0 |
0 |
0 |
0 |
23 |
3 |
0 |
0 |
0 |
0 |
0 |
23 |
4 |
1 |
0 |
1 |
1 |
2 |
30 |
5 |
1 |
1 |
1 |
1 |
6 |
32 |
6 |
1 |
1 |
1 |
1 |
8 |
35 |
7 |
1 |
1 |
3 |
2 |
11 |
35 |
8 |
1 |
1 |
4 |
3 |
11 |
36 |
9 |
3 |
1 |
5 |
4 |
14 |
36 |
10 |
6 |
4 |
8 |
5 |
16 |
40 |
11 |
6 |
6 |
11 |
7 |
18 |
40 |
12 |
6 |
7 |
13 |
9 |
18 |
40 |
13 |
6 |
9 |
14 |
9 |
18 |
40 |
14 |
6 |
9 |
14 |
9 |
18 |
40 |
15 |
7 |
9 |
16 |
11 |
18 |
40 |
16 |
7 |
9 |
16 |
13 |
18 |
40 |
17 |
7 |
9 |
17 |
15 |
20 |
40 |
18 |
7 |
9 |
17 |
16 |
21 |
40 |
19 |
7 |
9 |
17 |
16 |
21 |
40 |
20 |
7 |
9 |
18 |
18 |
22 |
40 |
21 |
7 |
9 |
18 |
18 |
22 |
40 |
% dead on day 21 |
17.5 |
22.5 |
45 |
45 |
55 |
100 |
At the start of the test 40 daphnids were introduced at each concentration.
Table 3. Cumulative number of juveniles produced by parental Daphnia in each control and test vessel
Measured conc. D-8 (mg/L) |
Cumulative no. of juveniles produced in each vessel |
|||||
8 days |
11 days |
13 days |
15 days |
18 days |
21 days |
|
Control |
32, 47, 49, 45 |
238, 192, 222, 219 |
287, 241, 258, 219 |
448, 395, 379, 365 |
698, 653, 614, 637 |
984, 1020, 831, 1028 |
Total |
173 |
871 |
1005 |
1587 |
2602 |
3863 |
0.139 |
60, 58, 35,76 |
215, 215, 216, 214 |
254, 262, 249, 302 |
422, 360, 406, 442 |
721, 542, 599, 699 |
892, 754, 844, 1006 |
Total |
229 |
860 |
1067 |
1630 |
2561 |
3496 |
0.492 |
89, 115, 81, 45 |
291, 271, 203, 211 |
291, 279, 216, 253 |
446, 450, 362, 382 |
673, 668, 591, 641 |
794, 739, 713, 813 |
Total |
330 |
976 |
1039 |
1640 |
2573 |
3059 |
1.45 |
23, 17, 10, 38 |
203, 160, 170, 180 |
254, 194, 269, 275 |
338, 296, 323, 343 |
534, 499, 491, 521 |
676, 725, 575, 649 |
Total |
88 |
713 |
992 |
1300 |
2045 |
2625 |
5.04 |
0, 0, 0, 0 |
49, 57, 91, 40 |
93, 74, 118, 58 |
154, 105, 144, 100 |
225, 144, 159, 153 |
262, 196, 167, 251 |
Total |
0 |
237 |
343 |
503 |
681 |
876 |
14.8 |
No juveniles were produced |
|||||
Total |
Table 4. Mean cumulative number of juveniles produced per adult in each vessel
Measured conc. D-8 (mg/L) |
Mean cumulative no. of juveniles produced per adult |
|||||
8 days |
11 days |
13 days |
15 days |
18 days |
21 days |
|
Control |
3.2, 4.7, 5.4, 4.5 |
23.8, 19.2, 24.6, 21.9 |
29.9, 24.6, 29.1, 21.9 |
50.0, 41.7, 44.2, 38.1 |
81.3, 70.4, 73.6, 72.1 |
117.1, 111.2, 100.7, 121.0 |
Mean |
4.5 |
22.4 |
26.4 |
43.5 |
74.4 |
112.5 |
0.139 |
6.0, 5.8, 3.9, 7.6 |
21.5, 21.5, 24.0, 21.4 |
26.4, 26.7, 28.7, 30.2 |
47.4, 40.7, 51.1, 45.8 |
84.8, 66.7, 78.7, 74.4 |
106.2, 97.0, 113.7, 108.5 |
Mean |
5.8 |
22.1 |
28.0 |
46.3 |
76.2 |
106.4 |
0.492 |
9.9, 11.5, 8.1, 4.5 |
35.2, 27.1, 20.3, 25.3 |
35.2, 28.1, 22.2, 30.6 |
66.2, 49.5, 46.5, 49.0 |
111.6, 85.8, 84.7, 86.0 |
135.8, 100.0, 105.0, 110.6 |
Mean |
8.5 |
27.0 |
29.0 |
52.8 |
92.0 |
112.9 |
1.45 |
2.6, 1.7, 1.0, 3.8 |
22.6, 19.6, 17.0, 18.0 |
28.3, 24.5, 28.0, 29.9 |
37.6, 39.1, 35.7, 38.4 |
59.4, 68.1, 69.3, 60.7 |
79.7, 100.4, 97.3, 79.0 |
Mean |
2.3 |
19.3 |
27.7 |
37.7 |
64.4 |
89.1 |
5.04 |
0, 0, 0, 0 |
6.1, 8.1, 11.4, 6.7 |
14.9, 10.9, 16.8, 9.7 |
27.1, 16.1, 22.0, 16.7 |
41.3, 22.6, 25.0, 25.5 |
50.6, 31.3, 27.7, 41.8 |
Mean |
0 |
8.1 |
13.1 |
20.5 |
28.6 |
37.9 |
14.8 |
No juveniles were produced |
Description of key information
D-8 was tested for 21-day juvenile production study in Daphnia magna according to OECD Guideline 202. The 21-day median lethal concentration (LC50) of D-8 to parental Daphnia was calculated to be 2.70 mg/L. The 21-day EC50 values for inhibition of reproduction were 1.90 mg/L based on the total number of juveniles produced at each mean measured concentration, and 2.94 mg/L based on the mean cumulative number of juveniles produced per adult. In conclusion, D-8 significantly affected both the survival of the parental generation and their ability to produce viable juveniles at concentrations above 0.139 mg/L. Therefore, NOEC was determined to be 0.139 mg/L.
Key value for chemical safety assessment
Fresh water invertebrates
Fresh water invertebrates
- Effect concentration:
- 0.139 mg/L
Additional information
The effect of D-8 on the survival and reproduction of Daphnia magna has been assessed under semi-static exposure conditions over a period of 21 days in accordance with OECD 202, Part 2 (1984). Groups of forty Daphnia were exposed to D-8 at nominal concentrations of 0.15, 0.5, 1.5, 5 and 15 mg/L, selected following a ten-day range finding test. A control group of Daphnia was exposed to dilution water alone. The test was conducted at 20 ± 0.7 °C in treated tap water of hardness 45 to 59 mg/L as CaCO3 at pH 7.1 to 8.2. Exposure levels were monitored by HPLC analysis. The numbers of mobile, immobile and dead parental Daphnia were recorded each day of the test, and the numbers of juveniles produced were counted three times each week.
In groups exposed to D-8, mortality of the parental Daphnia after 21 days ranged from 22.5 % at the lowest level (0.139 mg/L) to 100 % at the highest level (14.8 mg/L). Mortality of the parental control Daphnia after 21 days was 17.5 %.
The 21-day median lethal concentration (LC50) of D-8 to parental Daphnia was calculated to be 2.70 mg/L, with 95 % confidence limits of 1.80 - 3.65 mg/L.
The 21-day EC50 values for inhibition of reproduction were 1.90 mg/L, based on the total number of juveniles produced at each mean measured concentration, and 2.94 mg/L, based on the mean cumulative number of juveniles produced per adult. In conclusion, D-8 significantly affected both the survival of the parental generation and their ability to produce viable juveniles at concentrations above 0.139 mg/L. Therefore, NOEC was determined to be 0.139 mg/L.
In a supporting study, long-term toxicity tests of D-8 were performed using Daphnia magna. The Daphnia magna (21 d) chronic toxicity tests were conducted as outlined in Organization for Economic Cooperation and Development (OECD) test guidelines 211 (OECD, 2012). For D. magna chronic toxicity tests, 10 neonates per group were exposed to D-8 (0, 0.06, 0.12, 0.25, 0.5, 1 mg/L). The endpoints were survival of parent animal, first day of neonate production, number of neonates per parent animal, number of neonates per brood, and growth. In D. magna exposed to D-8 for 21 d, no adverse effects of survival and reproduction were observed up to the highest test concentration of 1 mg/L. However, the body length was significantly reduced when exposed to 1 mg/L of D-8. The NOEC value for reproduction of D-8 was 1 mg/L. D-8 was also tested for 7 days in Moina macrocopa, an indigenous species in Asia, in concentrations of 0, 0.01, 0.03, 0.11, 0.33, 1 mg/L. The endpoints were survival of parent animal, first day of neonate production, number of neonates per parent animal, number of neonates per brood. The first day of reproduction was significantly increased at 1 mg/L D-8. The number of young per female, one of the most sensitive endpoints related to the reproduction, was significantly decreased with treatment of 0.3 mg/L of D-8. The NOEC value for reproduction of D-8 was 0.1 mg/L.
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