Dodecyl(2-hydroxy-3-sulphonatopropyl)dimethylammonium

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:

key study

Study period:

October 5 2017 to November 1 2017

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:

April 13, 2004

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Chemical name: Reaction products of chloromethyloxirane, sodium bisulfite and dodecyldimethylamine, which consist of 2-hydroxy-4,4-dimethyl-4-azoniahexadecanesulfonate as a major component
Other name: A-20HD
CAS number: Main component: 13197-76-7
Di-ol: 35396-47-5
Disulfonate: 163709-39-5
Cation: 103947-09-7
Chlorohydrin: 126-83-0
Purity: 37.5% (test item concentration was corrected for its purity)
Impurity: Sodium chloride: 6.5%
Water: 56.0%
Lot number: 2361
Physicochemical properties:
Water solubility: ≥100 g/L
Melting point: -10°C
Appearance: Pale yellow clear liquid
Density: 1.108 g/cm3 (25°C)
Storage: Test item was stored in a cold and dark storage place
Identification and stability of test item under storage conditions: Infrared (IR) spectrum of the test item measured at the laboratory was confirmed to be identical to that provided by the sponsor. In addition, the IR spectrum of the test item after completion of the experiment was identical to that before the experiment initiation, indicating the test item was stable under storage conditions.

Analytical monitoring:

yes

Details on sampling:

Samples of test solutions (approximately 10 ml) were taken from all test levels for chemical analysis at the start and end of exposure to measure test item concentrations. Another solution was sampled separately from the preparation container for measurement at the start of exposure. Equal volumes of solution were sampled from the middle layer of the test solution in test vessels in each test level at the end of exposure.

Vehicle:

no

Details on test solutions:

Dechlorinated tap water aerated sufficiently and temperature controlled was used as dilution water. Chemical characteristics of the dilution water met the standards provided with the standard operation procedure of the testing laboratory.

Test sample (0.4 g) was weighed and dissolved in dilution water (1.5 L) to prepare 100 mg/L stock solution (correcting by the purity (37.5%) of the test item). A control consisting of only dilution water was included. In addition, NaCl (0.0519 g) was weighed and dissolved in dilution water (3 L) to prepare NaCl dilution water of 17.3 mg/L. Necessary volumes of stock solution and NaCl dilution water were mixed and stirred in a preparing container to prepare each test concentration. In the 100 mg/L and NaCl control, the stock, the stock solution and NaCl dilution water were each divided in test vessels. Four replicates per test level were inlcuded in the study design.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test organism source: The University of Sheffield, UK
Date of supply: July 9, 1990
Acclimation: Young daphnids produced by parents cultured in the testing laboratory were used in experiments. The Parents used to obtain young daphnids were bred in the same quality of water (dechlorinated tap water), water temperature (20±1°C) and photoperiod (16:8 h light:dark) used in testing. Parent animals used for the test were from the same lot and their age and survival rate was 14-days old and 95%, respectively. Chlorella vulgaris at 0.1-0.2 mgC (Organic carbon content)/day per daphnid was fed to the parents once per day.
Age at test initiation: <24-h old
Test group allocation: Random sampling
Feeding: Test organisms were not fed during testing

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

None

Post exposure observation period:

Not applicable

Hardness:

32 mg/L

Test temperature:

20.0 to 20.1°C

pH:

7.8 to 7.9

Dissolved oxygen:

8.5 to 8.9 mg/L

Salinity:

Not applicable (freshwater test)

Conductivity:

13 mS/m

Nominal and measured concentrations:

Nominal concentration: 0 (dilution water control), 9.53, 17.1, 30.9, 55.6, 100 mg/L, a NaCl control was also included (17.3 mg/L)

Details on test conditions:

Tests were conducted in 100 ml glass beakers covered with a transparent plastic lid and placed in a plastic tank water bath (Warming/cooling unit; Type HCA 250 (Sato craft)). Each test vessel contained 100 ml of test solution, with 20 daphnids were used per test level (5 daphnids/test vessel). Test organisms were not fed throughout testing. The temperature of test solution was maintained at 20±1°C, dissolved oxygen concentration was >3 mg/L without aeration, pH was not adjusted and test vessels were kept under a 16:8 h (light:dark) photoperiod. Dissolved oxygen concentration, pH and temperature were measured at the start of exposure from a solution separate from the preparation container and at the end of exposure from one test vessel in each test level. Dissolved oxygen concentration was measured via dissolved oxygen meter (HQ30d (HACH)), pH was measured via pH meter (HM-21P (DKK-TOA)) and temperature was measured via thermometer (of glass stick type). The appearance of test solution was observed at the start and end of exposure.

Test organisms were observed for immobility and other symptoms at 24 and 48 hours of exposure. Daphnids were considered immobile when they were unable to swim within 15 seconds after gentle agitation of the test vessel.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

71.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% confidence limits: 60.1-88.0 mg/L

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

36.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% confidence limits: 30.9-43.3 mg/L

Details on results:

The minimum concentration causing 100% immobility at 48 hours was 100 mg/L. Maximum concentration causing no immobility at 48 hours was 9.53 mg/L. Observed abnormal responses during exposure were immobilisation, lethargic and reduced activity. Control organisms showed 0% immobility during exposures and no abnormal responses thereby meeting the study validity criterion. No adverse effect was shown in the NaCl control, thus it was concluded that the observed effects on test organisms was caused by the test item.

All test solutions were colourless and clear at the start and end of exposure. Measured dissolved oxygen concentration values met the study validity criterion. Measured concentrations of the test item in test solutions at the start of exposure were 9.71-104 mg/L (99.3-102% of nominal concentration). The measured concentrations of the test item were kept within ±20% of the nominal concentration.

Results with reference substance (positive control):

A 48-h acute immobilisation test with the reference substance, potassium dichromate, with the test organisms was periodically conducted at the testing laboratory. The most recent results before the current study (4 to 6 April 2017) resulted in a 48-h EC50 of 0.15 mg/L. This value was within the stipulated range (mean ±2S.D.) [mean ± S.D.: 0.23±0.06 mg/L (n=100)] to background data in this laboratory.

Reported statistics and error estimates:

The EC50 was calculated via Probit analysis via a computer program (running on Microsoft Excel software) developed by the testing laboratory and the 95% confidence limits and slope of the regression line at 95% confidence limits calculated. Study results were estimated based on nominal concentrations since measured test item concentrations in solution were maintained within ±20% of the nominal concentrations during exposure. The 24-h and 48-h EC50 values of the test item for Daphnia magna were 71.4 mg/L (95% confidence limits: 60.1-88.0 mg/L) and 36.6 mg/L (95% confidence limits: 30.9-43.3 mg/L), respectively.

Immobility

Nominal concentration (mg/L)	Vessel	24 hours		48 hours
		Number of immobilised daphnids / Total daphnids	Immobility (%)	Number of immobilised daphnids / Total daphnids	Immobility (%)
Control	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
NaCl control	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
9.53	A	0 / 5	0	0 / 5	0
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
17.1	A	0 / 5	0	1 / 5	5
	B	0 / 5		0 / 5
	C	0 / 5		0 / 5
	D	0 / 5		0 / 5
30.9	A	0 / 5	0	1 / 5	30
	B	0 / 5		2 / 5
	C	0 / 5		1 / 5
	D	0 / 5		2 / 5
55.6	A	2 / 5	35	5 / 5	85
	B	1 / 5		4 / 5
	C	2 / 5		4 / 5
	D	2 / 5		4 / 5
100	A	5 / 5	75	5 / 5	100
	B	3 / 5		5 / 5
	C	4 / 5		5 / 5
	D	3 / 5		5 / 5

 

Validity criteria fulfilled:

yes

Conclusions:

The 24-h and 48-h EC50 values of A-20HD were estimated to be 71.4 (95% confidence limit: 60.1-88.0 mg/L) and 36.6 mg/L (95% confidence limit: 30.9-43.3 mg/L), respectively. Test item concentration was corrected for its purity. No adverse effect was shown in the NaCl control, thus it was concluded that the observed effects on test organisms was caused by the test item.

Executive summary:

The acute toxicity of A-20HD to Daphnia magna was determined in a static study according to the OECD 202 Guideline (‘Daphnia sp., Acute immobilisation Test’). The study was conducted to confirm the effect of the test item on the test organisms below the solubility of the test item in dilution water. In order to confirm the effect of NaCl as an impurity in the test sample, the NaCl concentration in all test concentrations was made to the same concentration (17.3 mg/L) as the maximum test concentration, and a NaCl control (17.3 mg/L as NaCl concentration) was separately set.

 

Test organisms were exposed to nominal test concentrations of 0 (dilution water control), 9.53, 17.1, 30.9, 55.6, 100 mg/L for 48 hours. The test item is composed of the main component and five sub-components, therefore when quantitatively measuring test concentrations, each component was individually quantified and the total of these defined as the test item concentration. Chemical analysis confirmed that the nominal concentrations of the test item in solution were maintained within ±20% of the nominal concentrations during exposure. The biological results of the study were therefore related to nominal concentrations of the test item.

 

The 48-h EC50 was calculated to be 36.6 mg/L (95% confidence limit: 30.9-43.3 mg/L). Test item concentration was corrected for its purity. No adverse effect was shown in the NaCl control. It was therefore concluded that the observed effects on test organisms was caused by the test item.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

May 1995

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

No chemical analysis included in study design

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984

Deviations:

yes

Remarks:

The reported study is a screening study, not conducted according to GLP and no chemical analysis performed. The screening report contains no information on culture conditions, use of reference substances etc.

GLP compliance:

no

Specific details on test material used for the study:

Betadet S-20
Batch number 7244

The Certificate of Analysis (not included in the report) provides the following information:
Batch: 7244 (29/03/1995)
Appearance: Liquid, transparent, slightly yellowish, fluid
Dry matter: 44.5%
pH: 6.8
Sodium chloride: 6.4%

The principal constituent (included in the 44.5% 'dry matter') represented approximately 30% (w/w) of the tested batch.

Analytical monitoring:

no

Details on sampling:

Not applicable

Vehicle:

no

Details on test solutions:

The test material was directly dispersed in water and a100 mg/L stock solution was prepared. This stock was diluted with reconstituted water to prepare the test media.

Test organisms (species):

Daphnia magna

Details on test organisms:

No details reported.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

None

Post exposure observation period:

Not applicable

Hardness:

270 mg CaCO3/L (theoretical)

Test temperature:

21°C

pH:

7.6 - 8.0 (test medium comprising reconstituted water)

Dissolved oxygen:

Not reported

Salinity:

Not applicable (freshwater test)

Conductivity:

<5 micro Sieverts/cm (test medium comprising reconstituted water)

Nominal and measured concentrations:

Nominal exposure concentrations, 0 (control), 0.1, 0.6, 1.0, 3.0, 6.0, 10 and 30 mg/L.

Details on test conditions:

No details reported

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

4.8 mg/L

Nominal / measured:

nominal

Conc. based on:

other: principal constituent

Basis for effect:

mobility

Remarks on result:

other: Calculated from reported endpoint as (16.0 x 0.3), based on 30% (w/w) principal constituent content of tested batch.

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

16 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 12 - 22 mg/L.

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 30 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

30 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

The 24h EC50 of Betadet S-20 was observed to be >30 mg/L (the highest nominal concentration tested) and the 48 h EC50 was calculated to be 16 mg/L with a corresponding NOEC of 6.0 mg/L.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Analysis was performed according to the method described by Thompson, 1947 (Thompson, WR, 1947, Bact. Reviews (11, 115-145)

Daphnia observations

Nominal concentration (mg/L)	Cumulative immobilised Daphnia (initial population = 20)
	24 hours		48 hours
	Total	%	Total	%
Control	0	0	0	0
0.1	0	0	0	0
0.6	0	0	0	0
1.0	0	0	0	0
3.0	0	0	0	0
6.0	0	0	0	0
10	0	0	10	50
30	0	0	14	70

 

Validity criteria fulfilled:

not specified

Remarks:

Immobilisation after 48 hours in the control group was <10%, but water quality parameters were not reported.

Conclusions:

The 24h EC50 of Betadet S-20 was observed to be >30 mg/L (the highest nominal concentration tested) and the 48 h EC50 was calculated to be 16 mg/L with a corresponding NOEC of 6.0 mg/L. The 48 h EC50 is equivalent to 4.8 mg principal constituent/L after correction for the 30% (w/w) p.c. content of the tested batch of Betadet S-20.

Executive summary:

The acute toxicity of Betadet S-20 to Daphnia magna was determined in a static study carried out according to OECD test guideline 202 (1984). Daphnia magna were exposed to nominal test concentrations of 0 (control), 0.1, 0.3, 1.0, 3.0, 6.0, 10 and 30 mg/L for 48 hours. No analytical verification was performed therefore the results of the study are based in the nominal exposure concentrations. 

The 24h EC50 of Betadet S-20 was observed to be >30 mg/L (the highest nominal concentration tested) and the 48 h EC50 was calculated to be 16 mg/L with a corresponding NOEC of 6.0 mg/L. The 48 h EC50 and NOEC are equivalent to 4.8 and 1.8 mg principal constituent/L, based on the 30% (w/w) p.c. content of the tested batch of Betadet S-20.

Description of key information

The key short-term toxicity to aquatic invertebrates study (Adachi, 2017) resulted in 24-h and 48-h EC50 values of 71.4 and 36.6 mg/L, respectively for Daphnia magna exposed to A-20HD. Test item concentration was corrected for its purity. The study met all of the validity criteria prescribed by the standard OECD 202 Guideline and is considered to be reliable.

 

A supporting study (Wetton, 1995) resulted in a 24-h EC50 of Betadet S-20 of >30 mg/L (the highest nominal concentration tested) and a 48-h EC50 of 16 mg/L, with a corresponding NOEC of 6.0 mg/L. The 48 h EC50 equates to 4.8 mg principal constituent/L, based on the 30% (w/w) p.c. content of the tested batch of Betadet S-20. However, given that no chemical analysis of test concentrations was included, this study is considered unreliable.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

36.6 mg/L