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

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jan 6, 1993 - Feb 10, 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: US EPA TSCA, 40 CFR, Part 797.1600

Deviations:

no

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 in the dark
- Other: Test material was supplied to the laboratory as a series of aqueous dispersions collectively referenced as SS0140.01R. Low radiolabelled dispersions were supplied as 0.30 % (SS0190.01R), 0.63% (SS0188.01R), 1.2% (SS0187.01R), 2.6% (SS0185.01R), and 4.9% (SS0184.01R) MDEA-Esterquat by weight. High radiolabelled dispersions were supplied as 0.30% (SS0189.01R), 1.2% (SS0186.01R), and and 5.5% (SS0183.01R) MDEA-Esterquat by weight. All dispersions were prepared by adding the MDEA-Esterquat test material (SS0134.01) and a labelled tracer (SS0132.01R) 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 -1, 0, 1, 7, 14, 21, 28 and 35. Samples for speciation were taken from the 0.686, 2.68 and 9.76 mg/L exposure concentrations on study days -1, 0, 1, 14, 28 and 35.
- 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 pH adjusted deionized water (pH 2.5, concentrated HCl). Stock solutions were delivered via syringe pumps to the dilutor. Test solutions were delivered to the system for 10 days prior to test initiation. Low-activity radiolabelled stock solutions were replaced by high-activity radiolabelled stock solutions periodically for the 0.686, 2.68 and 9.76 mg/L exposure concentrations for analysis of MDEA-Esterquat and hydrolysis components by RAD-TLC and LSC.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Cloudiness in the 9.76 mg/L exposure. Cloudiness eventually in the 5.30 mg/L exposure. Brown precipitate in the control and all exposure concentrations attributed to Perche Creek dilution water.

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead Minnow
- Source: In-house breeding colony

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs): 5 females and 5 males
- Method of collection of fertilised eggs: Removed from tiles by rolling under water
- Subsequent handling of eggs: All eggs pooled, impartially added to exposure chambers not more than two at one time

POST-HATCH FEEDING
- Start date: Day 4
- Type/source of feed: Days 4-11, fed Brachionus sp. (Aquaculture Supply, FL, USA) and Artemia sp. (Aquarium Products, MD, USA); Days 12-15, fed Artemia sp.; Days 16-33, fed Artemia sp. and Salmon Starter mash (Zeigler Bros., PA, USA); Days 24-33, also fed Number 1 granule Salmon Starter (Zeigler Bros., PA, USA). Days 34-35, not fed.
- Amount given: All chambers received same amount of food during each feeding. Sufficient food was added to allow for at least 30 minutes of feeding.
- Frequency of feeding: Three times per day Monday thru Friday; two to three times per day Saturday and Sunday.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

35 d

Hardness:

Control: mean 206 mg/L, SD 26.0 mg/L, range 160-230 mg/L
0.686 mg/L: mean 206 mg/L, SD 23.0 mg/L, range 160-228 mg/L
9.76 mg/L: mean 204 mg/L, SD 25.5 mg/L, range 158-226 mg/L

Test temperature:

Control: mean 25.2°C, SD 0.42°C, range 24.6-25.9°C
0.686 mg/L: mean 25.2°C, SD 0.36°C, range 24.6-25.8°C
1.41 mg/L: mean 25.1°C, SD 0.26°C, range 24.6-25.4°C
2.68 mg/L: mean 25.1°C, SD 0.33°C, range 24.6-25.7°C
5.30 mg/L: mean 25.1°C, SD 0.31°C, range 24.6-25.6°C
9.76 mg/L: mean 25.2°C, SD 0.53°C, range 24.3-26.1°C

pH:

Control: mean 8.0, SD 0.17, range 7.8-8.3
0.686 mg/L: mean 8.0, SD 0.18, range 7.8-8.3
9.76 mg/L: mean 8.0, SD 0.20, range 7.7-8.3

Dissolved oxygen:

Control: mean 7.0 mg/L, SD 0.79 mg/L, range 5.8-7.9 mg/L
0.686 mg/L: mean 7.0 mg/L, SD 0.84 mg/L, range 5.6-7.9 mg/L
1.41 mg/L: mean 7.0 mg/L, SD 0.80 mg/L, range 5.8-7.9 mg/L
2.68 mg/L: mean 7.0 mg/L, SD 0.68 mg/L, range 6.1-7.9 mg/L
5.30 mg/L: mean 6.8 mg/L, SD 0.64 mg/L, range 5.8-7.6 mg/L
9.76 mg/L: mean 6.4 mg/L, SD 0.52 mg/L, range 5.7-7.2 mg/L

Nominal and measured concentrations:

Nominal: 0, 0.625, 1.25, 2.5, 5.0, 10 mg/L
Measured: 0, 0.686, 1.41, 2.68, 5.30, 9.76 mg/L

Details on test conditions:

TEST SYSTEM
- Embryo cups (if used, type/material, size, fill volume): glass, 9 cm diameter, 40 mesh Nitex screen on bottom, oscillated vertically on rocker arm
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 21.7 cm x 15.3 cm x 14.3 cm, 4.7 L chamber volume
- Type of flow-through (e.g. peristaltic or proportional diluter): 0.5 L proportional dilutor
- Renewal rate of test solution (frequency/flow rate): 33 L/day/replicate (approx. 7 test chambers/day)
- No. of fertilized eggs/embryos per vessel: 25
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- Biomass loading rate: 0.021-0.10 g/L/day at study termination

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Perche Creek river water. Water was passed through a degassing system reservoir prior to filtration and UV sterilization
- Total organic carbon: 3-4.6 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-detected in quarterly water samples
- Alkalinity: 102-144 mg/L
- Conductivity: 374-535 µS/cm
- Intervals of water quality measurement: Temperature, DO, pH, conductivity, hardness, alkalinity on days 0, 1, 7 and every 7 days thereafter. Temperature and DO taken in control and each test level on scheduled days. Additional DO and temperature measurements were taken on days 17, 23, 25, 26, 32, 33. Conductivity, pH, hardness, alkalinity taken in Control, low and high test concentrations. Temperature continuously monitored in one chamber.
- Suspended solids: 24 mg/L in dilution water; 16.4 +/- 6.1 mg/L in control; 20 +/- 5.6 mg/L in low concentration; 36 +/- 17.5 in high concentration

OTHER TEST CONDITIONS
- Photoperiod: 16:8 light:dark
- Light intensity: 388-625 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Hatchability, pre-fry-reduction survival, post-fry-reduction survival, standard length, wet weight

VEHICLE CONTROL PERFORMED: no vehicle used

RANGE-FINDING STUDY
- Test concentrations: 1, 10, 50 mg/L (nominal)
- Results used to determine the conditions for the definitive study: survival LC50 12 mg/L at 25 days (21 days post hatch); weight and length reduction at 10 mg/L

POST-HATCH DETAILS
- Begin of post-hatch period: day 5
- No. of hatched eggs (alevins)/treatment released to the test chamber: 15
- Release of alevins from incubation cups to test chamber on day no.: 5

Reference substance (positive control):

no

Duration:

5 d

Dose descriptor:

NOEC

Effect conc.:

> 9.76 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

number hatched

Duration:

5 d

Dose descriptor:

NOEC

Effect conc.:

> 9.76 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

other: pre-fry reduction mortality

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

0.686 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

length

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

0.686 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

weight

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

0.686 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

other: post-fry reduction mortality

Duration:

35 d

Dose descriptor:

LOEC

Effect conc.:

1.41 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

other: post-fry reduction mortality

Duration:

35 d

Dose descriptor:

LC50

Effect conc.:

1.67 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: total activity by LSC

Basis for effect:

other: post-fry reduction mortality

Remarks on result:

other: 95% CL 1.45 and 1.91 mg/L

Details on results:

- Mortality/survival at embryo and larval stages: embryo stage survival at 0, 0.686, 1.41, 2.68, 5.30, 9.76 mg/L was 80.0, 88.0, 90.7, 89.3, 84.4, 64.9 %; larval stage survival at 0, 0.686, 1.41, 2.68, 5.30, 9.76 mg/L was 80.0, 91.1, 33.3, 53.3, 0, 22.9%.
- Overall mortality/survival: Not reported
- Days to hatch and numbers hatched: 2-5 days to hatch; % Hatch at 0, 0.686, 1.41, 2.68, 5.30, 9.76 mg/L was 80.0, 88.0, 90.7, 89.3, 87.0, 75.7%.
- Data for length and weight of surviving fish: lengths at 0, 0.686, 1.41, 2.68, 9.76 mg/L were 24.4, 24.4, 20.8, 22.8, 19.4 mm (no survival at 5.30 mg/L); weights at 0, 0.686, 1.41, 2.68, 9.76 mg/L were 0.234, 0.232, 0.148, 0.192, 0.103 g (no survival at 5.30 mg/L).
- Type of and number with morphological abnormalities: 1 fish in both 0 and 2.68 mg/L concentrations extremely small
- Type of and number with behavioural abnormalities: erratic swimming pattern, quiescence, loss of equilibrium, and surfacing observed in control and all exposure concentrations during the test, but involved only a few fish at any given time.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Hatchability, Pre-fry-reduction mortality and Post-fry-reduction mortality were analyzed using frequency analysis coupled with the chi-square statistic and Fisher's exact test.
LC50 values and the 95% confidence limits were calculated using a computerized program developed by Stephan et al. using the moving average test.
Growth data was assessed by a one-way ANOVA. A Dunnett's one-tailed means comparison procedure was used to determine those exposure concentrations exhibiting responses significantly less than the control. Histograms and a Shapiro-Wilks normality test were used to assess normality. Statistical significance at p

Validity criteria fulfilled:

yes

Remarks:

The NOEC of 0.686 mg/L met validity criteria of survival in each chamber. However, larval survival at test termination not concentration dependent. Survival in one control chamber prior to fry reduction and one control chamber at test termination <70% .

Conclusions:

The 35-day chronic toxicity of MDEA-Esterquat C16-18 and C18 unsatd. to early life stages of Fathead minnow, Pimephales promelas, was studied under flow-through conditions according to US EPA TSCA, 40 CFR, Part 797.1600. The 35-day NOEC values, based on mortality (post-fry reduction) and sub-lethal effects (length and weight) were 0.686 mg/L. The 5-day NOEC based on number hatched was >9.76 mg/L (measured concentrations each).

Executive summary:

The 35-day chronic toxicity of MDEA-Esterquat C16-18 and C18 unsatd. to early life stages of Fathead minnow, Pimephales promelas, was studied under flow-through conditions using river water, according to US EPA TSCA, 40 CFR, Part 797.1600. Fertililized eggs (25 eggs, reduced to 15 fry after hatch) were exposed to a control and measured concentrations of 0.686, 1.41, 2.68, 5.30 and 9.76 mg/L as total activity by LSC. The test system was maintained at 25 +/- 1°C and a pH of 8.0 (7.7 to 8.3). The 35-day NOEC values, based on mortality (post-fry reduction) and sub-lethal effects (length and weight) were 0.686 mg/L. The 5-day NOEC based on number hatched was >9.76 mg/L. Although the post-fry reduction survival did not monotonically decrease with exposure concentration, a NOEC of 0.686 mg/L could statistically be determined. Although survival in one control chamber prior to fry reduction and one control chamber at test termination was <70%, the NOEC of 0.686 mg/L met validity criteria of survival in each chamber of >70%.

This toxicity study is classified as reliable with restriction and satisfies the guideline requirement for early life toxicity study with fish.

Description of key information

35-day NOEC based on mortality and sub-lethal effects: 0.686 mg/L
5-day NOEC based on number hatched: >9.76 mg/L (effective) 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.686 mg/L

Additional information

The 35-day chronic toxicity of MDEA-Esterquat C16-18 and C18 unsatd. to early life stages of Fathead minnow, Pimephales promelas, was studied under flow-through conditions (using river water) according to US EPA TSCA, 40 CFR, Part 797.1600. Fertililized eggs (25 eggs, reduced to 15 fry after hatch) were exposed to a control and measured concentrations of 0.686, 1.41, 2.68, 5.30 and 9.76 mg/L as total activity by LSC. The test system was maintained at 25 +/- 1°C and a pH of 8.0 (7.7 to 8.3). The 35-day NOEC values, based on mortality (post-fry reduction) and sub-lethal effects (length and weight) were 0.686 mg/L. The 5-day NOEC based on number hatched was >9.76 mg/L. Although the post-fry reduction survival did not monotonically decrease with exposure concentration, a NOEC of 0.686 mg/L could statistically be determined. Although survival in one control chamber prior to fry reduction and one control chamber at test termination was <70%, the NOEC of 0.686 mg/L met validity criteria of survival in each chamber of >70%.