Quaternary ammonium compounds, di-C8-10-alkyldimethyl, chlorides

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

other: US FIFRA Guideline 72-2 Static Acute Toxicity Test with Daphnids

Deviations:

yes

Remarks:

Minor deviations from the protocol were not considered to have affected the results.

Principles of method if other than guideline:

Minor deviations from the protocol included (i) a slight modification to the food items (algae and trout food instead of algae and yeast); (ii) water temperature measured between 19 and 22 degrees C during the study, but the protocol required 20 to 22 degrees C; (iii) 10 daphnids per vessel were used in the definitive test instead of 5 daphnids stated in the protocol. The deviations were not considered to have affected the results.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Radio-labelled and non-labelled substance used
Radio-labelled 14C-DDAC is a clear liquid and >99% pure with a specific activity of 9.40 mCi/mmol
Non-labelled DDAC is a light yellow liquid containing 41.2% active ingrediant

Analytical monitoring:

yes

Details on sampling:

Samples were removed and analysed at 0, 24 and 48 hours from one replicate of each test concentration and the control. Samples were taken from the mid-point of the container with a volumetric pipette. Three QC samples were also prepared at each sampling interval.

Vehicle:

no

Details on test solutions:

Fortified well water was used for dilution to achieve mean measured test concentrations of 0, 19, 31, 51, 74 and 120 ug AI/L. The dilution water was the same source as the culture water for daphnids.

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnids were cultured within the Springborn Laboratory and maintained in fortified well water. The culture received a regulated photoperiod of 16 hours light and 8 hours dark. daphnids were fed 2.0 ml of the green algae Ankistrodesmus falcatus and 0.5ml of trout food once daily. The food solution contained 5 mg/ml trout food and approximately 40,000,000 cells/ml of algae. The ambient air temperature in the cutlure area was maintained at 20 +/- 2 degrees C.

Twenty daphnids <24 hours old were distributed to each concentration vessel (ten daphnids per replicate) within 30 minutes after the test solution had been prepared. Daphnids were not fed during the experiment.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

Test substance concentrations were measured at 0, 24 and 48 hour intervals.

Post exposure observation period:

No information reported

Hardness:

Total hardness as CaCO3 = 170 mg/L

Test temperature:

19 to 22 degrees C

pH:

8.2

Dissolved oxygen:

Dissolved oxygen saturation during the test ranged from 8.3 to 9.3 mg/L (or 89 to 102%) across the test concentrations and 7.3 to 9.1 (80 to 100%) saturation in the control samples.

Salinity:

No information reported

Conductivity:

Specific conductivity = 500 umhos/cm

Nominal and measured concentrations:

Nominal concentrations were 0, 13, 22, 37, 60 and 100 ug AI/L and mean measured concentrations were 0, 19, 31, 51, 74 and 120 ug AI/L.

A 1.0 mg AI/ml stock solution was prepared by diltuing 0.0213 g of non-labelled DDAC with 3.8 ml and 0.350 mg AI/ml radio-labelled 14C-DDAC stock solution and brought to volume with distille dwater in a 10 ml volumetric flask. Test solutions were stirred using a magnetic stirrer and teflon-coated stir bars for 30 seconds before being divided into two replicate vessels each containing 1000 ml..

Details on test conditions:

The tests were run in glass beakers and not aerated during the 48 hour study.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

94 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

74 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

Prior to the definitive test, a range-finding test was undertaken by the laboratory. In the range-finding study, daphnids were exposed under static conditions from 100 to 0.001 mg AI/l. 100% immobilisation was observed at the four highest concentrations tested (100, 10, 1.0 and 0.1 mg AI/L). No adverse effects were observed in the two lowest concentrations tested (0.01 and 0.001 mg AI/L). Based on these results, the nominal concentrations of 100, 60, 37, 22, 13 and 0 ug AI/L were selected for the definitive study.

The water quality parameters were unaffected by DDAC during the definitive study. Mean measured concentrations of the test substance were generally consistent between sampling intervals and followed the expected concentration gradient.

Results with reference substance (positive control):

No information reported

Reported statistics and error estimates:

The EC50 and 95% confidence intervals were calculated in a computer programme of C Stephan (Peltier, 1985). Three statistical methods were available including moving average angle analysis, probit analysis and non-linear interpolation with 95% confidence intervals calculated by binomial probability. Moving average angle and probit analysis only yield statistically sound results if at least two concentrations produce immobilisation of between 0 and 100% in the test population. The results are expressed using non-linear interpolation.

Validity criteria fulfilled:

yes

Conclusions:

In a 48-hour static acute toxicity study of Daphnia magna exposed to the test substance (DDAC or didecyldimethyl ammonium chloride) resulted in an EC50 of 94 ug AI/L (or 0.094 mg AI/L) as a measured concentration.

Executive summary:

An acute toxicity study of the test substance DDAC exposed to Daphnia magna in a static test resulted in a 48-hour EC50 of 0.094 mg AI/L. The study aligns with the OECD Test Guideline 202 and was undertaken by a commercial laboratory using its own daphnid culture. A range-finding test preceded the definitive test. No adverse effects was observed in the control groups of organisms.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to US EPA Office of Pesticide Programs guideline.

Qualifier:

according to guideline

Guideline:

EPA OPP 72-3 (Estuarine/Marine Fish, Mollusk, or Shrimp Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples of test solutions from each concentration and controls were be taken at the initiation (0 hour) and test termination (96 hour) in order to verify concentrations of radioactivity.

Vehicle:

no

Details on test solutions:

For analytical purposes, DDACarbonate was spiked with a small quantity of 14C-DDA Chloride.

Test organisms (species):

Americamysis bahia (previous name: Mysidopsis bahia)

Details on test organisms:

TEST ORGANISM
- Common name: Mysid Shrimp
- Strain: Mysidopsis Bahia
- Food type: Live brine shrimp, Artemia salina nauplii
- Frequency: Twice daily

ACCLIMATION
- Acclimation period: Not applicable

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Not applicable

Test temperature:

25 - 26 °C

pH:

7.4 - 7.9

Dissolved oxygen:

3.8 - 6.7 mg/L

Salinity:

29 - 31 ‰

Nominal and measured concentrations:

Nominal concentrations: 0, 13, 22, 35, 60 and 100 µg AI/L
Mean measured concentrations: 0, 12, 20, 30, 54 and 89 µg AI/L

Details on test conditions:

Test vessels were 1.9 L glass beakers containing 1 L of test solution. The test solution had a depth of 3.5 cm with a surface area of 280 cm2.
Groups of ten mysids (two replicates per treatment level and control) were exposed in a static system for 96 hours to nominal concentrations of 13, 22, 35, 60 and 100 µg AI/L. Dilution water controls were included. For analytical purposes, the test item was spiked with a small quantity of 14C-DDAC. Test vessels were maintained at 25 +/-1 °C and the area was illuminated with Durotest Vitalite® and General Electric Coolwhite® fluorescent lights, Light intensity was measured at the surface of the test solutions to be 70 footcandles (750 lux) and the photoperiod was 16 hours light and 8 hours darkness. Water samples were analysed radiometrically and the measured test concentrations were defined as 12, 20, 30, 54 and 890 µg/L. During the course of the study, water quality was monitored and daily observations for visible abnormalities and mortality were made and recorded.

Reference substance (positive control):

not required

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.073 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Remarks on result:

other: 0.054 to 0.089 mg AI/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.03 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

Following 96 hours of exposure, 90 % mortality was observed among Mysid shrimp exposed to the highest treatment level, 89 µg AI/L. No mortality was observed in the remaining treatment levels. The obtained 96-hour LC50 was 73 µg AI/L and the NOEC 30 µg AI/L. All the surviving mysids in the 89 µg AI/L treatment group were observed to be lethargic while one of the surviving mysids in the 54 µg AI/L group was noted to be lethargic. No sublethal effects (e.g. lethargy, loss of equilibrium) were observed among Mysid shrimp exposed to 12, 20 and 30 µg AI/L.
Although this study was conducted on a 45.9 % solution of the test material, a correction factor was used and the results given are for active ingredient.

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 value was estimated by non-linear interpolation to be 73 µg AI/L (95 % confidence interval calculated by binomial probability of 0.054 to 0.089 mg AI/L. The No-Observed-Effect Concentration established for this study was determined to be 30 µg AI/L.

Executive summary:

A study was carried out according to EPA OPP 72-3 (Estuarine/Marine Fish, Mollusk, or Shrimp Acute Toxicity Test). In a static test Mysid shrimp (Mysidopsis Bahia) were exposed to measured concentrations of 0, 12, 20, 30, 54 and 89 µg AI/L. The 96-hour LC50 value was estimated by non-linear interpolation to be 73 µg AI/L. The NOEC established for this study was determined to be 30 µg AI/L.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study according to US EPA Office of Pesticide Programs guideline.

Qualifier:

according to guideline

Guideline:

EPA OPP 72-2 (Aquatic Invertebrate Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples of test solutions from each concentration and controls were be taken at the initiation (0 hour) and test termination (48 hour) in order to verify concentrations of radioactivity.

Vehicle:

no

Details on test solutions:

For analytical purposes, DDACarbonate was spiked with a small quantity of 14C-DDA Chloride.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Daphnia magna
- Source: Daphnia were obtained from in-house cultures maintained by the laboratory
- Age at study initiation: less than 24 hours old
- Food type: combination a trout food suspension and a unicellular green algae
- Amount: 5 mg/mL trout food and 4.0E07 cells/mL of algae
- Frequency: once daily

ACCLIMATION
- Acclimation period: NA

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

168 mg CaCO3/L

Test temperature:

20 °C

pH:

3.6 - 8.7

Dissolved oxygen:

8.1 - 8.5 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal concentrations: 0, 13, 22, 36, 60 and 100 µg AI/L
Mean measured concentrations: 0, 13, 24, 33, 64 and 100 µg AI/L

Details on test conditions:

Test vessels were 1.6 L glass beakers containing 1 L of test solution. The test solution had a depth of 19 cm with a surface area of 90.3 cm2.
Groups of ten daphnids (two replicates per treatment level and control) were exposed in a static system for 48 hours to nominal concentrations of 13, 22, 36, 60 and 100 µg AI/L. Dilution water controls were included. For analytical purposes, the test item was spiked with a small quantity of 14C-DDAC. The daphnid culture area received a regulated photoperiod of 16 hours of light and 8 hours of darkness. Light at an intensity of 20 to 100 footcandles at the surface of the culture solutions was provided by DuroTest VitaLife® fluorescent bulbs. Water quality characteristics were measured at 0, 24 and 48 hours. Test solutions were prepared by diluting a stock solution of 14C-DDAC with non-radiolabelled test item and dilution water. The number of immobilised daphnids in each replicate test vessel was recorded at 24 and 48 hours of exposure.

Reference substance (positive control):

not required

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.066 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 0.033 to 0.100 mg AI/L

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 0.1 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.033 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

All organisms exposed to the lowest exposure level (13 µg/L) were immobilised within the initial 24 hours. Water quality measurements performed during this study established that, during this period, the pH of the exposure solutions of the lowest exposure level decreased to ca. 3.6. Analyses of the lowest exposure level demonstrated that measured concentrations were in agreement with the nominal fortified levels. No mortalities or sublethal effects were observed at the 24-hour interval among organisms exposed to the 24 and 33 µg/L treatment levels, indicating that the immobilisation observed at 13 µg/L was in fact due to low pH. Following 48 hours of exposure, immobilisation of 100 % and 45 % was observed among daphnids exposed to 100 and 64 µg/L respectively. The obtained 48-hour EC50 was 66 µg/L and the NOEC 33 µg/L. Sublethal effects were observed among several daphnids exposed to the 64 µg/L treatment level. No mortalities or sublethal effects were noted in the 24 and 33 µg/L treatment groups or in the control after 48 hours.
Although this study was conducted on a 45.9 % solution of the test material, a correction factor was used and the results given are for active ingredient.

Validity criteria fulfilled:

yes

Conclusions:

The 48-hour EC50 value was estimated by non-linear interpolation to be 66 µg AI/L (95 % confidence interval calculated by binomial probability of 33 to 100 µg AI/L. The No-Observed-Effect Concentration established for this study was determined to be 33 µg AI/L.

Executive summary:

A study was carried out according to EPA OPP 72-2 (Aquatic Invertebrate Acute Toxicity Test). In a static test Daphnia magna were exposed to nominal concentrations of 13, 22, 36, 60 and 100 µg AI/L and measured concentrations of 0, 13, 24, 33, 64 and 100 µg AI/L. The 48-hour EC50 value was estimated by non-linear interpolation to be 66 µg AI/L. The NOEC established for this study was determined to be 33 µg AI/L.

Description of key information

Two reliable acute toxicity study with the aquatic invertebrate Daphnia magna, and using the read across substance, N,N-Didecyl-N,N-dimethylammonium chloride

and DDACarbonate respectively are available with a 48-hour LC50 of 0.094 mg/L and 0.066 mg/L respectively.

The LC50 of 0.066 mg/L will be taken as the short-term toxicity in aquatic invertebrates endpoint for C8 -10DAQ.

The 48-hour EC50 was determined to be 0.094 mg/L. The key study exposed Daphnia magna to DDACarbonate for 48-hours

and resulted in an EC50 of 0.066 mg/l (the most sensitive endpoint value)

(Springborn Laboratory, 1994

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.1 mg/L