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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-08-10 to 2017-11-16
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:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Periodic analyses of water for potential contaminants were not performed according to GLP Standards, but were performed using a certified laboratory and standard US EPA analytical methods.
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
Daphnia magna
Details on test organisms:
Test Organism
The cladoceran, Daphnia magna, was selected as the test species for this study. This species is representative of an important group of aquatic invertebrates and was selected for use in the test based upon past history of use in the laboratory. Daphnids used in the test were <24-hour old neonates obtained from cultures maintained by EAG Laboratories in Easton, Maryland. Identification of the species was verified by the supplier of the original stock culture.

Adult daphnids were cultured in water from the same source and at approximately the same temperature as used during the test. During the 2-week period immediately preceding the test, water temperatures in the cultures ranged from 19.6 to 20.4ºC, the pH of the water ranged from 8.0 to 8.5, and the dissolved oxygen concentrations were ¿7.6 mg/L (¿84% of saturation). The four adult daphnids used to supply neonates for the test were held for at least 21 days prior to collection of the juveniles for testing, and had each produced at least one previous brood. Adult daphnids in the culture had produced an average of at least three young per adult per day over the 7-day period prior to the test. The adults showed no signs of disease or stress, no ephippia were produced during the holding period, and mortality in the culture stock was 0% in the two-day period prior to test initiation.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Test temperature:
19.5 - 20.5 °C
pH:
8.3 - 8.6
Dissolved oxygen:
>= 8.0 mg/L (>= 89% ASV): no aeration
Nominal and measured concentrations:
Nominal concentrations: 0 (Control), 6.3, 13, 25, 50, 100 mg/L
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
not determinable
Remarks:
Maximum observed effect under 50%

RESULTS AND DISCUSSION

Observations and Measurements

Measurements of temperature, pH and dissolved oxygen of the water in the test chambers are presented below:

Table 1Temperature, Dissolved Oxygen and pH of Water in the Test Chambers

 

Nominal

WAF

Loading Rate

(mg a.i./L)

 

 

 

Replicate

0 Hour

 

24 Hours

 

48 Hours

Temp.1

(°C)

DO2

(mg/L)

 

pH

 

Temp.1

(°C)

DO2

(mg/L)

 

pH

 

Temp.1

(°C)

DO2

(mg/L)

 

pH

Negative

A

20.1

8.5

8.3

 

19.6

8.4

8.5

 

19.7

8.3

8.6

Control

B

20.2

8.5

8.4

 

19.6

8.4

8.5

 

19.6

8.3

8.6

 

C

20.2

8.5

8.4

 

19.6

8.4

8.5

 

19.6

8.4

8.6

 

D

20.2

8.5

8.4

 

19.5

8.4

8.5

 

19.6

8.4

8.6

 

 

 

 

 

 

 

 

 

 

 

 

 

6.3

A

20.3

8.5

8.3

 

19.6

8.4

8.5

 

19.6

8.4

8.6

 

B

20.3

8.4

8.3

 

19.6

8.4

8.5

 

19.7

8.4

8.6

 

C

20.3

8.4

8.3

 

19.6

8.4

8.5

 

19.7

8.4

8.6

 

D

20.3

8.4

8.3

 

19.6

8.4

8.5

 

19.6

8.4

8.6

 

 

 

 

 

 

 

 

 

 

 

 

 

13

A

20.3

8.3

8.3

 

19.6

8.4

8.5

 

19.7

8.4

8.6

 

B

20.3

8.3

8.3

 

19.6

8.5

8.5

 

19.7

8.5

8.6

 

C

20.3

8.4

8.3

 

19.6

8.5

8.5

 

19.7

8.6

8.6

 

D

20.3

8.4

8.3

 

19.6

8.5

8.5

 

19.6

8.6

8.6

 

 

 

 

 

 

 

 

 

 

 

 

 

25

A

20.4

8.2

8.3

 

19.6

8.5

8.5

 

19.7

8.6

8.6

 

B

20.4

8.1

8.3

 

19.6

8.4

8.5

 

19.7

8.6

8.6

 

C

20.5

8.2

8.3

 

19.6

8.5

8.5

 

19.7

8.6

8.6

 

D

20.4

8.1

8.3

 

19.6

8.5

8.5

 

19.7

8.6

8.6

1  Manual temperature measurements. Temperature monitored continuously during the test ranged from 19.50 to 19.68°C, measured to the nearest 0.01°C.

2  A dissolved oxygen concentration of 5.4 mg/L represents 60% saturation at 20°C in freshwater.

Table 1 (Continued)Temperature, Dissolved Oxygen and pH of Water in the Test Chambers

 

Nominal

WAF

Loading Rate

(mg a.i./L)

 

 

 

Replicate

0 Hour

 

24 Hours

 

48 Hours

Temp.1

(°C)

DO2

(mg/L)

 

pH

 

Temp.1

(°C)

DO2

(mg/L)

 

pH

 

Temp.1

(°C)

DO2

(mg/L)

 

pH

50

A

20.5

8.0

8.3

 

19.6

8.6

8.5

 

19.7

8.6

8.6

 

B

20.5

8.0

8.3

 

19.6

8.7

8.5

 

19.7

8.5

8.6

 

C

20.4

8.0

8.3

 

19.6

8.6

8.5

 

19.7

8.6

8.6

 

D

20.4

8.1

8.3

 

19.6

8.6

8.5

 

19.7

8.6

8.6

 

 

 

 

 

 

 

 

 

 

 

 

 

100

A

20.4

8.1

8.3

 

19.6

8.6

8.5

 

19.7

8.6

8.6

 

B

20.3

8.1

8.3

 

19.6

8.6

8.5

 

19.7

8.6

8.6

 

C

20.3

8.0

8.3

 

19.6

8.6

8.5

 

19.6

8.6

8.6

 

D

20.3

8.0

8.3

 

19.6

8.6

8.5

 

19.7

8.6

8.6

1  Manual temperature measurements. Temperature monitored continuously during the test ranged from 19.50 to 19.68°C, measured to the nearest 0.01°C.

2  A dissolved oxygen concentration of 5.4 mg/L represents 60% saturation at 20°C in freshwater.

Water temperatures were within the 20 ± 1°C range established for the test. Measurements of pH ranged from 8.3 to 8.6. Dissolved oxygen concentrations remained =8.0 mg/L (=89% of saturation) throughout the test. The measurements of hardness, alkalinity, specific conductance and TOC in the dilution water at the beginning of the test were typical of EAG Laboratories-Easton well water (see Table below).

 

Table 2Specific Conductance, Hardness, Alkalinity and Total Organic Carbon (TOC)

in the Dilution Water at Test Initiation

 

Parameter

Day 0

 

Specific Conductance

384

 

     (µS/cm)

 

 

 

 

 

Hardness

144

 

     (mg/L as CaCO3)

 

 

 

 

 

Alkalinity

176

 

     (mg/L as CaCO3)

 

 

 

 

 

Total Organic Carbon (TOC)

<2

 

     (mg C/L)

 

 


Light intensity at the beginning of the test was 651 lux at the surface of the water of one representative test chamber. Test solutions in the test chambers at these nominal WAF loading rates appeared clear and colorless during the test, with no evidence of precipitation observed.

Daily observations of immobility and other signs of toxicity observed during the test are presented below:

Table 3 Cumulative Immobility and Observations

 

Nominal

WAF

Loading Rate

(mg a.i./L)

 

 

 

Rep.

 

 

No.

Exposed

~2.5 Hours

 

24 Hours

 

48 Hours

 

Cumulative

Percent

Immobility

Number

Immobile1

 

Observations2

 

Number

Immobile1

 

Observations2

 

Number

Immobile1

 

Observations2

Negative

A

5

0

5 Q,AN

 

0

5 Q,AN

 

0

5 AN

0

Control

B

5

0

4 Q,AN; 1 AN

 

0

4 Q,AN; 1 AN

 

0

2 Q,AN; 3 AN

 

 

C

5

0

4 Q,AN; 1 AN

 

0

1 Q,AN; 4 AN

 

0

1 Q,AN; 4 AN

 

 

D

5

0

5 AN

 

0

5 AN

 

0

5 AN

 

 

 

 

 

 

 

 

 

 

 

 

 

6.3

A

5

0

5 Q,AN

 

0

1 Q,AN; 4 AN

 

0

1 Q,AN; 4 AN

0

 

B

5

0

5 Q,AN

 

0

2 Q,AN; 3 AN

 

0

5 AN

 

 

C

5

0

5 AN

 

0

5 AN

 

0

5 AN

 

 

D

5

0

1 Q,AN; 4 AN

 

0

5 AN

 

0

5 AN

 

 

 

 

 

 

 

 

 

 

 

 

 

13

A

5

0

5 Q,AN

 

0

5 Q,AN

 

0

2 Q,AN; 3 AN

0

 

B

5

0

5 Q,AN

 

0

5 Q,AN

 

0

3 Q,AN; 2 AN

 

 

C

5

0

5 Q,AN

 

0

2 Q,AN; 3 AN

 

0

2 Q,AN; 3 AN

 

 

D

5

0

5 AN

 

0

5 AN

 

0

5 AN

 

 

 

 

 

 

 

 

 

 

 

 

 

25

A

5

0

5 Q,AN

 

0

3 Q,AN; 2 AN

 

0

3 C; 2 AN

5

 

B

5

0

5 Q,AN

 

0

5 AN

 

0

1 Q,AN; 4 AN

 

 

C

5

0

3 Q,AN; 2 AN

 

0

2 Q,AN; 3 AN

 

1

2 C; 2 AN

 

 

D

5

0

5 Q,AN

 

0

3 Q,AN; 2 AN

 

0

1 Q,C; 3 Q,AN; 1 AN

 

1  Cumulative number of immobile daphnids. Any immobile daphnids were removed from the test chambers at each observation interval.

2  Observations of surviving organisms: AN = appear normal; C = lethargy; Q,AN = trapped at water surface but appear normal after gentle submersion; Q,C = trapped at water surface and appear lethargic after gentle submersion.

 

Table 3 (Continued) Cumulative Immobility and Observations

 

Nominal

WAF

Loading Rate

(mg a.i./L)

 

 

 

Rep.

 

 

No.

Exposed

~2.5 Hours

 

24 Hours

 

48 Hours

 

Cumulative

Percent

Immobility

Number

Immobile1

 

Observations2

 

Number

Immobile1

 

Observations2

 

Number

Immobile1

 

Observations2

50

A

5

0

5 Q,AN

 

0

1 Q,C; 3 Q,AN; 1 AN

 

0

1 Q,AN; 4 C

0

 

B

5

0

5 Q,AN

 

0

1 Q,C; 2 Q,AN; 2 AN

 

0

1 C; 1 Q,C; 2 Q,AN; 1 AN

 

 

C

5

0

5 Q,AN

 

0

2 Q,AN; 3 AN

 

0

2 C; 2 Q,AN; 1 AN

 

 

D

5

0

4 Q,AN; 1 AN

 

0

1 Q,C; 1 Q,AN; 3 AN

 

0

1 C; 2 Q,AN; 2 AN

 

 

 

 

 

 

 

 

 

 

 

 

 

100

A

5

0

5 Q,AN

 

0

1 Q,C; 4 AN

 

0

1 Q,C; 1 Q,AN; 3 AN

0

 

B

5

0

4 Q,AN; 1 AN

 

0

2 Q,AN; 3 AN

 

0

5 AN

 

 

C

5

0

4 Q,AN; 1 AN

 

0

2 Q,AN; 3 AN

 

0

1 C; 2 Q,C; 2 AN

 

 

D

5

0

4 Q,AN; 1 AN

 

0

1 Q,AN; 4 AN

 

0

1 Q,C; 4 AN

 

1  Cumulative number of immobile daphnids. Any immobile daphnids were removed from the test chambers at each observation interval.

2  Observations of surviving organisms: AN = appear normal; C = lethargy; Q,AN = trapped at water surface but appear normal after gentle submersion; Q,C = trapped at water surface and appear lethargic after gentle submersion.

In the negative control and all treatment groups, floating daphnids were observed but appeared normal after gentle submersion and was not considered treatment related. All daphnids in the negative control group, including those floating daphnids described above, appeared normal throughout the test. All daphnids in the 6.3 and 13 mg a.i./L treatment groups, including those floating daphnids described above also appeared normal throughout the test, with no immobile daphnids or overt signs of toxicity observed. Percent immobility in the 6.3, 13, 25, 50 and 100 mg a.i./L treatment groups at test termination was 0, 0, 5, 0 and 0%, respectively. Signs of toxicity observed among the surviving daphnids in the 25, 50 and 100 mg a.i./L treatment groups at test termination included lethargy.

Therefore in this study, the no-immobility level was 100 mg a.i./L and the NOEL was 13 mg a.i./L. EL50 values at 24 and 48 hours were determined from the immobility data and are shown in the table below.

Table 4 EL50ValuesBased on Nominal WAF Loading Rates

 

 

Time

EL501

(mg a.i./L)

95% Confidence Interval

(mg a.i./L)

Statistical

Method

24 Hours

> 100

--2

NA3

48 Hours

> 100

--2

NA3

1 The EL50value is the theoretical WAF loading rate that produced 50% immobility.

2 95% confidence limits could not be calculated with the immobility data obtained.

3 NA = not applicable; <50% immobility precluded statistical calculation of an EL50value.

 

Conditions for the Validity of the Test

The following criteria were used to judge the validity of the test and were met in this study:

1. Immobility of the daphnids in the control group will not exceed 10% by the end of the test. In this study, immobility in the control group was 0%.

2. The dissolved oxygen concentration will be at least 60% of the air-saturation value throughout the test. In this study, the dissolved oxygen concentration remained =89% of saturation.

 


Validity criteria fulfilled:
yes
Conclusions:
CONCLUSIONS
The cladoceran, Daphnia magna, was exposed for 48 hours under static conditions to five nominal water accommodated fraction (WAF) loading rates of 1,12-Dodecanediol Bismethacrylate (CAS #72829-09-5) ranging from 6.3 to 100 mg a.i./L. Based on the nominal WAF loading rates, the 48-hour EL50 value (the theoretical WAF loading rate that produced 50% immobility) was >100 mg a.i./L, the highest loading rate tested. The no-immobility level was 100 mg a.i./L and the NOEL was 13 mg a.i./L.
Executive summary:

The cladoceran, Daphnia magna, was exposed for 48 hours under static conditions to five nominal water accommodated fraction (WAF) loading rates of 1,12-Dodecanediol Bismethacrylate (CAS #72829-09-5) ranging from 6.3 to 100 mg a.i./L.

Based on the nominal WAF loading rates, the 48-hour EL50 value (the theoretical WAF loading rate that produced 50% immobility) was >100 mg a.i./L, the highest loading rate tested. The no-immobility level was 100 mg a.i./L and the NOEL was 13 mg a.i./L.

Description of key information

A study assessing the acute toxicity of the test item to Daphnia magna was conducted in accordance with the OECD 202 and OPPTS 850.1010 Test Guidelines and GLP requirements.

As the maximum observed effect was below 50%, the 48h-EL50 was not mathematically determinable and it was considered to be greater than the highest loading rate tested: 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
100 mg/L

Additional information