Dimethyldioctylammonium chloride

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:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Standard 48-hour Daphnia study carried out by a commercial laboratory, including an initial range finding study.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

The study preceded standardised intenrational protocols, but is carried out by a commercial laboratory to the standards of the time.

Deviations:

not specified

Principles of method if other than guideline:

Following a range finding test, young Daphnia magna were exposed for 48 hours to a range of five concentrations of the test substance with 20 organisms per vessel. The organisms were fed and maintained at 21 degrees C with observations of survival made at 6, 24 and 48 hours.

GLP compliance:

no

Specific details on test material used for the study:

The test substance is referred to by its product name "Bardac LF". No further information is provided.

Analytical monitoring:

not specified

Details on sampling:

No information available

Vehicle:

no

Details on test solutions:

A range finding test was initially undertaken using 5 daphnids per test vessel. The range was found by beginning at 1ppm (1 mg/L) and dcreasing the amount of the test material by a factor of 10 until a range between no deaths and 100 percent of deaths was established. For the full test, the test material was dispensed into the test vessels in the form of 0.1% (w/v) solution in water. The test vessels were 250 ml glass beakers containing 200 ml of water.

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna were 12 +/- 12 hours old and obtained from Cannon Laboratories daphnid culture.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

The number of surviving individuals were recorded for each concentration at 6, 24 and 48 hours.

Post exposure observation period:

No information reported

Hardness:

No information reported

Test temperature:

21 +/- 2 degrees C

pH:

No information reported

Dissolved oxygen:

No information reported, but aeration was discontinued during the test to avoid loss of the test material

Salinity:

No information reported

Conductivity:

No information reported

Nominal and measured concentrations:

No information reported

Details on test conditions:

Test concentrations were 0, 0.075, 0.1, 0.18, 0.28, and 0.42 mg/l. Daphnids were fed 0.2 ml of diluted mixture of trout starter and alfalfa meal daily.

Reference substance (positive control):

not specified

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.24 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

LC50 determination was calculated according to Litchfield, J T Jr and Wilcoxon F (1949). A simplified method of evaluating dose-effect experiments. J. Pharm. and Exp. Therap. 96, 99-113.

Results with reference substance (positive control):

No information reported

Reported statistics and error estimates:

LC50 = 0.24 mg/l (0.20 to 0.29 mg/l) with confidence limits at P = 0.05.

No effects on survival were seen in the negative control and at the lowest concentration tested (0.075 mg/l). An LC20 was reported at 0.42 mg/l.

Validity criteria fulfilled:

not specified

Conclusions:

In a 48-hour static survival study of Daphnia magna exposed to the test substance resulted in an LC50 of 0.24 mg/l (nominal concentration).

Executive summary:

An acute toxicity study of the test substance C8DAQ exposed to Daphnia magna in a static test resulted in a 48-hour LC50 of 0.24 mg/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 survival test. No mortality 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

Study period:

14 to 16 November 2017 (experiemental study)

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)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch no.of test material: 6021-038
- Expiration date of the lot/batch: 23/05/2019
- Purity: 95.72%
- Manufacture date: 23/05/2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient temperature, protected from light
- Stability under test conditions: No data
- Solubility and stability of the test substance in the solvent/vehicle: No data
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Blank control and 0.0625, 0.125, 0.250, 0.500 and 1.00 mg/L (nominal)
- Sampling method: 0h samples were taken after preparation of the test concentrations; 48h samples were taken from test vessels
- Sample storage conditions before analysis: All samples stored at room temperature until the start of analysis, prepared samples were stored in the autosampler at room temperature until analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a nominal 1.00 mg/L stock solution was prepared in dilution water and test substance dissolved by manual agitation. Test concentrations were prepared by dilution of the nominal 1.00 mg/L stock with dilution water
- Eluate: Dilution water (Elendt M4) according to OECD 2002, Annex 3, 2004. Modified to total hardness 160-180 mg CaCOs/L
- Differential loading: No
- Controls: Blank control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Visually clear throughout exposure period

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Staus (Clone 5)
- Justification for species other than prescribed by test guideline: Not applicable
- Source: Institut fur Wasser, Boden und Lufthygiene, Berlin, Germany
- Age of parental stock (mean and range, SD): No data
- Feeding during test: No
- Food type: Not applicable
- Amount: Not applicable
- Frequency: Not applicable

ACCLIMATION
- Acclimation period: None

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

No data

Salinity:

No data

Conductivity:

No data

Nominal and measured concentrations:

Blank control and 0.0625, 0.125, 0.250, 0.500 and 1.00 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- Type (delete if not applicable): Loosely covered with watch glasses
- Material, size, headspace, fill volume: Glass beakers, 50 mL capacity, 20mL fill volume
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Static test , no test solution renewal
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): Not applicable

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 (according to OECD 202)
- Total organic carbon: No data
- Particulate matter: No data
- Metals: No data
- Pesticides: No data
- Chlorine: No data
- Alkalinity: No data
- Ca/mg ratio: No data
- Conductivity: No data
- Salinity: No data
- Culture medium different from test medium: No
- Intervals of water quality measurement: No data

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark
- Light intensity: diffuse light, 1500 lux maximum

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility every 24 hours

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

>= 0.025 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Nominal test concn (mg/L)	% immobilisation
	0 hour	48 hour
Control	0	0
0.0625	0	0
0.125	0	0
0.250	15	55
0.500	55	100
1.00	100	100

Validity criteria fulfilled:

yes

Conclusions:

The EC50, based on immobility, was determined to be 0.248 mg/L (95% CI 0.133 to 0.462 mg/L), based on nominal concentrations under the conditions of the test.

Executive summary:

In a 48-hour acute toxicity study the aquatic invertebrate,Daphnia magna, was exposed to a range of test concentrations under static conditions in accordance with the OECD test guideline 202 and in compliance with GLP. The nominal test concentrations were a control, 0.0625, 0.125, 0.250, 0.500 and 1.00 mg/L. Analytical measurement showed the test concentrations remained within ±20% of the nominal concentrations throughout the exposure period, therefore, results were calculated on the basis of nominal test concentrations.

Under the conditions of the test, the EC50 (immobility) was determined to be 0.248 mg/L (95% CI 0.133 to 0.462 mg/L), based on nominal concentrations.

This acute toxicity study is considered acceptable and satisfied the guideline requirements for a short-term toxicity study to aquatic invertebrates.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Standard 48-hour Daphnia magna study carried out by a commercial laboratory, including an initial range finding study.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

The test preceded international standardised protocols, but is similar to the OECD 202 Guideline

Deviations:

not specified

Principles of method if other than guideline:

An acute toxicity study with Daphnia magna exposed for 48-hours to the test substance, following an initial range finding study. The study was undertaken by a commercial laboratory to the standards of the time.

GLP compliance:

no

Specific details on test material used for the study:

Test substance = Bardac LF (Lot No. B3909)
Colour = clear yellow liquid
Source = Compound received on 22 April 1977 from the manufacturer
Odour = None
Total Hardness = 14.7 GPG (grains per gallon) (equivalent to 251.6 mg/L)
Iron = 0.7 mg/L
pH = 7.5
Conductivity (mmhos/cm) = 520
Nitrate = 1.7 mg/L
Silica = 7.0 mg/L
Total cations = 15.3 GPG (equivalent to 261.9 mg/L)
Total anions = 15.3 GPG (equivalent to 261.9 mg/L)

Analytical monitoring:

not specified

Details on sampling:

No information reported

Vehicle:

no

Details on test solutions:

The test material was dispensed into the bioassay vessles in the form of 0.10 percent (w/v) solution in water. The following concentration gradient was used: 0, 0.05, 0.075, 0.10, 0.18 and 0.28 mg/L.

Test organisms (species):

Daphnia magna

Details on test organisms:

The test organisms, Daphnia magna, were cultured at Cannon Laboratories. Young Daphnia were tested at 12 +/- 12 hours old.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

The number of surviving Daphnia was recorded at 6, 24 and 48 hours.

Post exposure observation period:

No information reported

Hardness:

251.6 mg/L

Test temperature:

18 +/- 1 degrees C

pH:

7.5

Dissolved oxygen:

No information reported

Salinity:

No information reported

Conductivity:

Conductivity (mmhos/cm) = 520

Nominal and measured concentrations:

Not specified, but taken as nominal concentrations.

Details on test conditions:

Twenty daphnids per test concentration were used. The test was run in 250 ml glass beakers containing 200 ml of water. Aeration was discontinued during the study to prevent the loss of test material. Feeding was also discontinued through the duration of the study. A photoperiod of 16 hours light was maintained during the study.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

LC50 determination was calculated according to Litchfield, J T Jr and Wilcoxon F (1949). A simplified method of evaluating dose-effect experiments. J. Pharm. and Exp. Therap. 96, 99-113.

Results with reference substance (positive control):

No information reported

Reported statistics and error estimates:

LC50 = 0.10 mg/L (0.085 to 0.12 mg/L) at the 95% confidence limits.

No mortality was observed in the negative controls.

Validity criteria fulfilled:

not specified

Conclusions:

In a 48-hour static survival study of Daphnia magna exposed to the test substance resulted in an LC50 of 0.10 mg/l (nominal concentration).

Executive summary:

An acute toxicity study of the test substance C8DAQ exposed to Daphnia magna in a static test resulted in a 48-hour LC50 of 0.10 mg/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 survival test. No mortality was observed in the control groups of organisms.

Reference 4

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

Justification for type of information:

Laboratory study carried out under GLP conditions to a standard United States FIFRA Guideline 72-2 protocol for an acute toxicity test with daphnids. Minor deviations were made to the protocol, such as exposing 10 daphnids per test concentration (from the 5 daphnids stated in the protocol) and a slightly amended food solution, but the laboratory does not consider these deviations to have affected the test results.

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.

Description of key information

The Key Study is a reliable acute toxicity test using the test substance dioctyldimethyl ammonium chloride (C8DAQ). The test substance was exposed to Daphnia magna in a static test and resulted in a 48-hour LC50 of 0.10 mg/l (Roberts, 1977). 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 survival test. No mortality was observed in the control groups of organisms. This study is selected as the Key Study for its reliability, its more conservative LC50 concentration compared to the Reinert (1976) study and the use of the test substance.

Six supporting studies are available, two for the test substance dioctyldimethyl ammonium chloride, C8DAQ, (Klix 2017 and Reinert, 1976), another for the read-across substance didecyldimethyl ammonium chloride, DDAC (LeLievre, 1990) and another three for the read-across substance N,N-Didecyl-N,N-dimethylammonium carbonate (DDACcarbonate), all by Springborn (2004) .

In a reliable study by Klix (2017), Daphnia magna were exposed to the test substance for 48 hours resulting in an LC50 of 0.248 mg/L. In a reliable study by Reinert (1976), Daphnia magna were exposed to the test substance for 48 hours which resulted in an LC50 of 0.24 mg/L.

In a reliable study by LeLievre (1990), using the read-across substance DDAC Daphnia magna were exposed to the read-across substance for 48 hours resulting in an EC50 based on immobilisation of 0.094 mg/L.

In three reliable studies all by Springborn (2004), on the read-across substance, DDACarbonate, a 96 hour LC50 for the mysid shrimp (Mysidopsis Bahia) was estimated by non-linear interpolation to be 73 µg AI/L, a 48-hour EC50 for Daphnia magna value was estimated by non-linear interpolation to be 66 µg AI/L and a 96 hour LC50 for the Eastern oyster (Crassostrea virginica) was estimated by non-linear interpolation to be 110 µg AI/L.

The results of all six studies are very close (0.066 to 0.248 mg/L) and demonstrate consistency in the concentrations at which Daphnia magna are adversely affected by the test substance and the read-across substances.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.1 mg/L