Ethanaminium, 2-hydroxy-N-(2-hydroxyethyl)-N,N-dimethyl-, esters with C16-18 and C18-unsatd. fatty acids, chlorides

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Feb 1, 1993 - Feb 22, 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1330 (Daphnid Chronic Toxicity Test)

Deviations:

yes

Remarks:

(TOC of 2.48-5.47 mg/L exceeded maximum allowable concentration of 2 mg/L)

GLP compliance:

yes

Remarks:

US EPA TSCA, 40 CFR, Part 792

Specific details on test material used for the study:

- Name of test material (as cited in study report): SS0140.01R (MDEA-Esterquat)
- Substance type: quaternized ammonium compound
- Storage condition of test material: room temperature
- Other: Test material was supplied to the laboratory as a series of aqeous dispersions collectively referenced as SS0140.01R. The dispersions were 0.125% (SS0203.01R), 0.25% (SS0202.01R), 0.5% (SS0201.01R), 1% (SS0200.01R), and 2% (SS0199.01R) MDEA-Esterquar by weight and were prepared by adding a MDEA-Esterquat stock compound (SS0134.01) and a labelled tracer (SS0130.01R) to pH 2.5 distilled water.
- Physical state: solid (SS0134.01)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Samples for LSC were taken from all exposure concentrations on study days -4, 0, 3, 8, 14 and 21. Samples for speciation were taken from the 0.25, 1.0 and 4.0 mg/L exosure concentrations on study days -4, 0, 3, 8, 14 and 21. On study day 21, a sample from the 2.0 mg/L exposure was taken due to mortality in the 4.0 mg/L exposure concentration.
- Sampling method: Samples were collected for the determination of MDEA-Esterquat and hydrolysis products (speciation) by LSC and RAD-TLC. Duplicate samples were taken for LSC analyses from each test and control chamber, added to a scintillation vial and scintillation cocktail was added. For RAD-TLC, triplicate samples were collected and adjusted to pH 2.5 with HCl .
- Sample storage conditions before analysis: Samples for RAD-TLC analyses were immediately extracted by SPE after the pH adjustment to 2.5.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solutions were prepared by diluting the dispersions with sterile deionized water. Stock solutions were delivered via syringe pumps to the dilutor. Test solutions were delivered to the system for ~6 days prior to test initiation.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: In-house culture obtained from the Columbia National Fisheries Research Laboratory
- Age of parental stock (mean and range, SD): 14 days old
- Feeding during test
- Food type: Algal suspension of Selenastrum capricornutum and Ankistrodesmus falcatus; suspension of trout chow and yeast.
- Amount: An equal volume of algal suspension that provided at least 2 x 10^8 cells/mL per chamber; 1 mL of 2.5 mg/L per test chamber of trout chow and yeast suspension.
- Frequency: Twice daily

ACCLIMATION
- Acclimation period: Parental stock 14 days old with 4 prior broods.
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: the algae species Selenastrum capricornutum and/or Ankistrodesmus falcatus; a supplement of trout chow and Fleishmann's active dry yeast (Saccharomyces sp.)
- Feeding frequency: daily

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

Control: 216-254 mg/L
0.27 mg/L: 218-236 mg/L
1.0 mg/L: 220-238 mg/L
2.0 mg/L: 236-242 mg/L
3.9 mg/L: 218-242 mg/L

Test temperature:

Control: 20-21°C C
0.27 mg/L: 20-21°C
1.0 mg/L: 20-21°C
2.0 mg/L: 21°C
3.9 mg/L: 20-21°C

pH:

Control: 8.1-8.4
0.27 mg/L: 8.2-8.4
1.0 mg/L: 8.2-8.4
2.0 mg/L: 8.3
3.9 mg/L: 8.2-8.4

Dissolved oxygen:

Control: 6.9-8.1 mg/L
0.27 mg/L: 7.0-8.1 mg/L
1.0 mg/L: 7.3-8.2 mg/L
2.0 mg/L: 7.4-7.6 mg/L
3.9 mg/L: 7.4-8.0 mg/L

Nominal and measured concentrations:

Nominal: 0, 0.25, 0.50, 1.0, 2.0, 4.0 mg/L
Measured: 0, 0.27, 0.47, 1.0, 2.0, 3.9 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 1 L chambers with 50-mesh stainless steel screens
- Type of flow-through (e.g. peristaltic or proportional diluter): 0.5 L proportional diluter
- Renewal rate of test solution (frequency/flow rate): 5.5 volume additions per day at an average rate of 3.9 mL/min to each test chamber.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: ~1 daphnid per 100 mL dilution water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: river water from Perche Creek near Columbia, Missouri. Dilution water was passed through a series of three ultraviolet sterilizers just prior to entering the dilutor. Each sterilizer contained a 100 micron filter.
- Total organic carbon: Day 0: 5.47 mg/L; Day 7: 2.69 mg/L; Day 21: 2.48 mg/L.
- Metals: Boron 0.070 mg/L, Iron <0.581 mg/L, Lead 0.0039 mg/L, Zinc 0.026 mg/L.
- Pesticides: non-detect in quarterly water samples
- Alkalinity: 128-152 mg/L
- Conductivity: 430-490 µmhos/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: Temperature, dissolved oxygen, pH, conductivity, hardness, and alkalinity were measured on days 0, 3, 7, 14 and 21 in the control, 0.27, 1.0 and 3.9 mg/L concentrations. Temperature of the water bath was recorded continuously.
- Suspended solids: Day 0: 7.0-18.5 mg/L; Day 7: 4-12.5 mg/L; Day 14: 14.5-19.5 mg/L; Day 21: 1.5-3.0 mg/L

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 h light:8 h dark
- Light intensity: 54 to 61 footcandles at the water surface

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Survival, abmormal effects and observance of first brood of the organisms were recorded daily. Reproductive success was measured by counting offspring in each concentration every Monday, Wednesday and Friday of the study.

VEHICLE CONTROL PERFORMED: no vehicle used

RANGE-FINDING STUDY
- Test concentrations: 0, 0.05, 0.10, 0.50, 1.0 and 5.0 mg/L.
- Results used to determine the conditions for the definitive study: A 10 day flow-through study was conducted with two reps of 10 organisms per concentration. After 10 days, 100% mortality occurred in the 5.0 mg/L exposure and 1 organism died in the 1.0 mg/L exposure. All levels with surviving daphnids reproduced on day 7 of the study except one 1.0 mg/L replicate which reproduced by day 8. All surviving daphnids appeared normal.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

1.7 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

mortality

Remarks on result:

other: 95% CL 1.5 and 1.9 mg/L

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

reproduction

Remarks:

(young/adult/reproduction day)

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

reproduction

Remarks:

(young/adult/reproduction day)

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

growth

Remarks:

(length and weight)

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

growth

Remarks:

(length and weight)

Details on results:

- Mortality of parent animals: mean percent survival in the control and mean measured concentrations of 0.27, 0.47, 1.0, 2.0 and 3.9 mg/L were 97.5 +/- 5.0%, 97.5 +/- 5.0%, 97.5 +/- 5.0%, 95.0 +/- 5.8%, 37.5 +/- 29%, 0.0 +/- 0.0%, respectively.
- No. of offspring produced per female: The mean and standard deviation of the number of young per adult per reproduction day in the control and mean measured concentrations of 0.27, 0.47, 1.0, and 2.0 mg/L were 13.06 +/- 1.12, 12.40 +/- 0.69, 11.76 +/- 1.60, 11.44 +/- 0.77 and 4.74 +/- 1.85. The total number of young for these concentrations were 7656, 7256, 6843, 6512 and 2233, respectively. There was no young per adult per reproduction day data for the mean measured concentration of 3.9 mg/L because no young were produced.
- Body length and weight of parent animals: The length of adult organisms in the control and mean measured concentrations of 0.27, 0.47, 1.0, and 2.0 mg/L were 4.85 +/- 0.07, 4.79 +/- 0.10, 4.69 +/- 0.11, 4.65 +/- 0.08 and 3.65 +/- 0.36 mm. There was no length data for the mean measured concentration of 3.9 mg/L because all the daphnids had died before the end of the study. The weight of adult organisms in the control and mean measured concentrations of 0.27, 0.47, 1.0, and 2.0 mg/L were 1.15 +/- 0.02, 1.25 +/- 0.09, 1.19 +/- 0.05, 1.18 +/- 0.04 and 0.29 +/- 0.12 mg. There was no weight data for the mean measured concentration of 3.9 mg/L because all the daphnids had died before the end of the study.
- Time to first brood release or time to hatch: Time to first brood in the control and mean measured concentrations of 0.27, 0.47, 1.0, and 2.0 mg/L wee all 7.0 +/- 0.0 days. There were no broods observed in the mean measured concentration of 3.9 mg/L.
- Other biological observations: Daphnids smaller than the control, tending to the bottom of the chamber, light discoloration, coated with extraneous materials and quiescent behavior were observed in the mean measured concentrations of 2.0 and 3.9 mg/L.

Reported statistics and error estimates:

Survival data were analyzed using frequency analysis that compared the test concentration to the control. This analysis was coupled with a one-tailed Fisher's exact test and the chi-square statistic. A one-way analysis of variance (ANOVA) and Dunnett's test were conducted to determine significant differences from the control. Reproduction, adult daphnid length and weight were assessed using an ANOVA and Dunnett's. A Sharpiro-Wilk test was computed within each test concentration to assess departures from a normal distribution. When a deviation from strict normality was indicated, results were examined for central tendency. Homogeneity of variance was also assessed. Statistical analyses were conducted using PC DOS SAS/STAT Release 6.03 with statistical significance at p

Validity criteria fulfilled:

yes

Remarks:

(<20% of control organisms immobilized, stressed or diseased; each control daphnid at test termination produced an average of >60 young)

Conclusions:

The 21-day chronic toxicity of MDEA-Esterquat C16-18 and C18 unsatd. to Daphnia magna was investigated under flow-through conditions according to EPA OTS 797.1330 (Daphnid Chronic Toxicity Test). The 21-day EC50 based on mortality was 1.7 mg/L (95% confidence limits of 1.5 and 1.9 mg/L). The 21 -day NOEC based on survival, number of young/adult/reproduction day, and growth (length and weight) was 1.0 mg/L. Production of offsprings in the treated groups indicated that MDEA-Esterquat C16-18 and C18 unsatd. had an effect on the reproduction at concentration greater than 1 mg/L.

Executive summary:

The 21-day chronic toxicity of MDEA-Esterquat C16-18 and C18 unsatd. to Daphnia magna was investigated under flow-through conditions (using river water) in a study conducted according to EPA OTS 797.1330 (Daphnid Chronic Toxicity Test). Daphnids were exposed to a control and test chemical at measured concentrations of 0, 0.27, 0.47, 1.0, 2.0 and 3.9 mg/L. The 21-day EC50 based on mortality was 1.7 mg/L (95% confidence limits of 1.5 and 1.9 mg/L). The 21 -day NOEC based on survival, number of young/adult/reproduction day, and growth (length and weight) was 1.0 mg/L. Production of offsprings in the treated groups indicated that MDEA-Esterquat C16-18 and C18 unsatd. had an effect on the reproduction at concentration greater than 1 mg/L.

This study is classified as reliable with restriction and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

Description of key information

Daphnia magna
21-day EC50 based on mortality: 1.7 mg/L (95% confidence limits of 1.5 and 1.9 mg/L effective)
21 -day NOEC based on survival, number of young/adult/reproduction day, and growth (length and weight): 1.0 mg/L
Production of offsprings in the treated groups indicated that MDEA-Esterquat had an effect on the reproduction at concentration greater than 1 mg/L.
This study is classified as reliable with restriction and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1 mg/L

Additional information

The 21-day chronic toxicity of MDEA-Esterquat C16-18 and C18 unsatd. to Daphnia magna was investigated under flow-through conditions (using river water) in a study conducted according to EPA OTS 797.1330 (Daphnid Chronic Toxicity Test). Daphnids were exposed to a control and test chemical at measured concentrations of 0, 0.27, 0.47, 1.0, 2.0 and 3.9 mg/L. The 21-day EC50 based on mortality was 1.7 mg/L (95% confidence limits of 1.5 and 1.9 mg/L). The 21-day NOEC based on survival, number of young/adult/reproduction day, and growth (length and weight) was 1.0 mg/L. Production of offspring in the treated groups indicated that MDEA-Esterquat C16-18 and C18 unsatd. had an effect on the reproduction at concentration greater than 1 mg/L.