Dimethyl[3-[(1-oxooctadecyl)amino]propyl][2-oxo-2-(tetradecyloxy)ethyl]ammonium chloride

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experimental start date was 12 Aug 2017, and the experimental completion date was 26 Oct 2017.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

Adopted April 13, 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Dimethyl[3-[(1-oxooctadecyl)amino]propyl][2-oxo-2-(tetradecyloxy)ethyl]ammonium chloride

EC Number:

272-964-2

EC Name:

Dimethyl[3-[(1-oxooctadecyl)amino]propyl][2-oxo-2-(tetradecyloxy)ethyl]ammonium chloride

Cas Number:

68921-83-5

Molecular formula:

C39H79N2O3.Cl

IUPAC Name:

dimethyl(3-octadecanamidopropyl)[2-oxo-2-(tetradecyloxy)ethyl]azanium chloride

Constituent 2

Reference substance name:

Propane-1,2-diol

EC Number:

200-338-0

EC Name:

Propane-1,2-diol

Cas Number:

57-55-6

Molecular formula:

C3H8O2

IUPAC Name:

propylene glycol

Test material form:

semi-solid (amorphous): gel

Details on test material:

Identification Quaternium-70
Appearance Amber gel
Batch 0002000146
Composition UVCB
Test item storage at room temperature
Stable under storage conditions until 02 April 2018 (retest date)

Specific details on test material used for the study:

Identification: Quaternium-70
Appearance: Amber gel
Batch 0002000146
Purity/Composition UVCB
Test item storage At room temperature
Stable under storage conditions until 02 April 2018 (retest date)
Test facility test item nr. 207517/A
Purity/correction factor Yes, correction factor is about 1/0.6 based on the fact that approx. 40% is propylene glycol, the correction factor is only to be used where applicable/possible
Test item handling No specific handling conditions required
Chemical name (IUPAC),
synonym or trade name: Ceraphyl 70

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Single samples for possible analysis were taken from all test concentrations and the control:
Frequency at t=0 h and t=48 h
Volume 2.0 mL from the approximate centre of the test vessels
Storage Not applicable, samples were transferred to the analytical laboratory at the Test Facility and analysed on the day of sampling.
At the end of the exposure period, the replicates were pooled at each concentration before sampling.

During the combined limit/range finding test, test samples were stored in the freezer (≤ -15°C).
On the day of analysis, the test samples were thawed at room temperature. The samples were diluted in a 1:1 (v:v) ratio with methanol and analyzed. If necessary, the samples were further diluted with 50/50 (v/v) methanol/ISO-medium to obtain concentrations within the alibration range.
During the final test, the samples were analyzed on the day of sampling and were diluted in a 1:1 (v:v) ratio with methanol and analyzed. If necessary, the samples were further diluted with 50/50 (v/v) methanol/ISO-medium to obtain concentrations within the calibration range.

Test solutions

Vehicle:

no

Details on test solutions:

The batch of Quaternium-70 tested was an amber gel and not completely soluble in test medium at the loading rates initially prepared. A correction was made for the purity/composition of the test item. A correction factor of 1.67 was used. Concentration/doses are expressed as mg Quaternium-70 per litre (mg Q-70/L).
In the combined limit/range-finding test, preparation of test solutions started with loading rates individually prepared at 1.7, 17 and 167 mg Q-70/L. A two-day period of slow magnetic stirring was applied to ensure maximum dissolution of the test item in medium. The obtained mixtures were allowed to settle over night. Thereafter, the aqueous Water Accommodated Fractions (WAFs) were collected by means of siphoning through glass wool and used as test concentrations. All test solutions were clear and colorless at the end of the preparation procedure, except for the highest test concentration, which was hazy.
In the range-finding and final tests, preparation of test solutions started with a stock solution of 100 mg Q-70/mL in acetone. No other treatment than vigorous shaking was needed to completely dissolve the test item in acetone. Stock solutions of lower concentrations were prepared by subsequent dilution of the highest stock concentration in acetone. Stock solutions of nominal 37 mg/mL and higher were clear and slightly yellow, while stock solutions of lower concentration were clear and colourless. Subsequently, 100 µL of each stock were spiked into 1 litre of continuously stirred medium. After an additional 15 minutes of magnetic stirring to ensure complete dissolution of the spiking solution in test medium, the obtained solutions were immediately used as test solutions in the range-finding test. In the final test, the test solutions were allowed to settle for one hour before they were used as test solutions. At the end of the preparation procedure, all test solutions were clear, colourless, and no Tyndall effect could be observed upon checking with a laser pen.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Source In-house laboratory culture with a known history.
Reason for selection This system has been selected as an internationally accepted invertebrate species.
Validity of batch Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Characteristics For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.
Breeding: start of each batch with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures is 4 weeks
Renewal of the cultures After 7 days of cultivation half of the medium twice a week.
Temperature of medium 18-22°C
Feeding Daily, a suspension of fresh water algae.
Medium M7, as prescribed by Dr. Elendt-Schneider (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).
The hardness 180 mg/L expressed as CaCO3 and the pH: 7.7 ± 0.3.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

20°C

pH:

8.0-8.2

Dissolved oxygen:

8.9-9.6 mg O2/L

Nominal and measured concentrations:

Nominal:0.22, 0.46, 1.0, 2.2 and 4.6 mg/L.
Stocks in acetone:
- concentrations measured according to the peak at m/z 485.5 were at 91-107% of nominal;
- concentrations measured based on the peak at m/z 623.6 were at 80-96% of nominal.
Initially measured test concentrations: 0.12, 0.21, 0.39, 0.81, 1.69 mg/L based on two selected peaks.
48-h average measured test concentrations: 0.026, 0.027, 0.029, 0.048, 0.066 mg/L based on two selected peaks.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.028 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Remarks:

m/z 485.5 and m/z 623.6

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.66 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The 48h-EC50for Daphnia magna exposed to Quaternium-70 was 0.028 mg Q-70/L based on average exposure concentrations (95% confidence interval between 0.028 and 0.029 mg Q-70/L).
The analytical system proved to be instable and responses of the calibration solutions at concentrations below 0.300 mg/L lacked repeatability at m/z 623.6. The limit of quantification was eventually set at 1.20 mg/L. The cause of the instability could not be determined and remained unknown.
In conclusion, the concentrations measured in the test solutions do not reliably represent actual exposure concentrations of the test item due to the limited quality and repeatability of the analytical measurements next to the complex nature of the test item. The complexity of the UVCB, together with its limited solubility and stability in water, make it highly unlikely that analytical support for determination of actual exposure concentrations of this test item can be improved.

Based on nominal concentrations, the 48h-EC50for Daphnia magna exposed to Quaternium-70 was 0.66 mg Q-70/L (95% confidence interval between 0.56 and 0.78 mg Q-70/L).

Results with reference substance (positive control):

The actual responses in the reference test with K2Cr2O7 are within the ranges of the expected responses at the different concentrations (i.e., the 48h-EC50 was between 0.3 and 1.0 mg/L).
Hence, the sensitivity of this batch of D. magna was in agreement with the historical data collected at Charles River Den Bosch.
The 24h-EC50 was 0.64 mg/L with a 95% confidence interval between 0.56 and 0.73 mg/L.
The 48h-EC50 was 0.48 mg/L with a 95% confidence interval between 0.43 and 0.53 mg/L.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the 48h-EC50 based on nominal concentrations for Daphnia magna exposed to Quaternium-70 was 0.66 mg Q-70/L (95% confidence interval between 0.56 and 0.78 mg Q-70/L).

Executive summary:

The objective of the study was to evaluate Quaternium-70 for its ability to generate acute toxic effects on the mobility ofDaphnia magnaduring an exposure period of 48 hours and, if possible, to determine the EC₅₀at 24 and 48 hours of exposure.

The study procedures described in this report were based on the OECD guideline No. 202, 2004.In addition, procedures were based on the test methods described in the OECD series on testing and assessment number 23, 2000.

The batch of Quaternium-70 tested was an amber gel and not completely soluble in test medium at the loading rates initially prepared. A correction was made for the purity/composition of the test item. A correction factor of 1.67 was used. Concentration/doses are expressed as mgQuaternium-70 per litre (mg Q-70/L).

A final test was performed based on the results of a preceding combined limit/range-finding and a range-finding test.

Preparation of test solutions started with a stock solution of 100 mg Q-70/mL in acetone. Stock solutions of lower concentrations were prepared by subsequent dilution of the highest stock concentration in acetone.Subsequently, 100 µL of each stock were spiked into 1 litre of continuously stirred medium. After 15 minutes of magnetic stirring to ensure complete dissolution of the spiking solution in test medium, the obtained solutions were allowed to settle for one hour before they were used as test solutions. At the end of the preparation procedure, all test solutions were clear, colourless, and no Tyndall effect could be observed upon checking with a laser pen.

Twenty daphnids per group (5 per replicate, quadruplicate) were exposed to an untreated control, a solvent control, and to nominal 0.22, 0.46, 1.0, 2.2 and 4.6 mg Q-70/L. The total exposure period was 48 hours and samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test.

No immobility was observed in the controls or the lowest test concentration throughout the test. At the end of the test, 5 and 95% of daphnids were immobile in nominal 0.46 and 1.0 mg Q-70/L, respectively, while all daphnids were found immobile in the two highest test concentrations.

Samples taken from all stock solutions and test concentrations were analysed employing two out of several peaks observed in the chromatograms of the test item.. All stock solutions measured were at 80-107% of the nominal concentration level according to either peak. All concentrations measured in the test solutions according to the peak atm/z 485.5 were at 5.8-43% of the nominal concentration level at the start of the test. These concentrations decreased below the limit of detection at the end of the test, except for the highest concentration which decreased to 6% of the initial concentration. The concentrations measured based on the peak atm/z 623.6 were below the limit of quantification at the start of the test, except the two highest test concentrations which were at 66-67% of the nominal concentration level. All concentrations decreased below the limit of quantification at the end of the test. Based on these results, the average exposure concentrations were calculated and used to determine the effect parameters. Additionally, the effect parameters were expressed based on nominal concentrations.

The study met the acceptability criteria prescribed by the study plan and was considered valid.

In conclusion, the 48h-EC₅₀for Daphnia magna exposed to Quaternium-70 was 0.028 mg Q-70/L based on average exposure concentrations (95% confidence interval between 0.028 and 0.029 mg Q-70/L). Based on nominal concentrations, the 48h-EC₅₀for Daphnia magna exposed to Quaternium-70 was 0.66 mg Q-70/L (95% confidence interval between 0.56 and 0.78 mg Q-70/L).