Ethanol, 2,2'-iminobis-, N-tallow alkyl derivs., N-oxides

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-03-04 to 2019-04-25

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:

13 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

A water accommodated fraction (WAF) equilibration trail was performed prior to exposure. The purpose of the equilibration trail is to determine the optimum mixing duration to be used for WAF preparation.
One WAF was prepared at each of three loading levels 1.0, 50.0 and 100 mg/L. At 2, 4, 6 and 24 hours after initiation of mixing, mixing was stopped and the solutions were allowed to settle for one hour. A sample of WAF was removed from each loading level and mixing was resumed.
The concentration of organic carbon that had solubilized into the WAF from the test substance was measured using the TOC analyzer (TOC Analyzer/TNML; Make: Shimadzu; Model: LCPH E200 ROHS).

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Strain: Straus
- Source: Environmental and Molecular Toxicology Division, Department of Zoology, Karnataka University, Karnataka, India
- Feeding during test: no
- Age of test animals at start of treatment: Less than 24 h old

ACCLIMATION
- Acclimation period: 48h
- Acclimation conditions: Same as test conditions
- Type and amount of food: Live yeast (Saccharomyces subcapitata) or algal cells (Pseudokirchneriella subcapitata) of 2 mL per liter

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not performed

Hardness:

Dose Range Finding study and Main study: 224 mg/L CaCO3

Test temperature:

Dose Range Finding Study: 19.9 - 21.5 °C
Main study: 20.0 - 21.1 °C

pH:

Dose Range Finding Study: 7.11 - 7.45
Main study: 7.13 - 7.53

Dissolved oxygen:

Dose Range Finding Study: 7.06 - 8.18 mg/L
Main study: 7.19 - 7.96 mg/L

Nominal and measured concentrations:

Nominal loading rates: 0.010, 0.021, 0.047, 0.103, 0.227 and 0.500 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Rectangular vessels
- Type: open
- Material, size: Glass, holding capacity of 100 mL
- Aeration during test: No
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: Reconstituted water with the pH 6.0 to 9.0, total hardness between 140 to 250 mg/L (as CaCO3). The dilution water was areated prior to use for the test so that the dissolved oxygen concentration has reached saturation.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light / 8 hours darkness

EFFECT PARAMETERS MEASURED: Mobility

RANGE-FINDING STUDY
- Test concentrations: 6 concentrations of 0.01, 0.1, 1.0, 10.0, 50.0 and 100.0 mg/L along with a negative control
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

0.082 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CI: 0.0696 - 0.0954

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

0.021 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

0.047 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

No clinical signs of toxicity or immobilization were observed in control and at the nominal loading rate of 0.010 mg/L during the 48 hours observation period.
Clinical signs of lethargy and daphnids localized on bottom of container was observed at 24 and 48 hours observation at the nominal loading rates of 0.021, 0.047, 0.103 and 0.227 mg/L.
Clinical signs of lethargy and daphnids localized on bottom of container was observed at 24 hours observation at the nominal loading rate of 0.500 mg/L.
The immobilization percentages of 0, 10, 35, 60, 80 and 100 % were observed at the nominal loading rates of 0.010, 0.021, 0.047, 0.103, 0.227 and 0.500 mg/L, respectively during the 48 hours observation period.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- 48h-EC50: 0.58 mg/L

Validity criteria of the test
- Immobilisation in the control group was 0 % at the termination of the test during main study (validity criterion: should not exceed 10 %).

- Dissolved oxygen concentration at the end of the test during main study was 7.96 mg/L (Validity criterion: should be ≥ 3 mg/L).

Table 1: Summary of clinical signs and immobilization during main study

Group	Nominal loading rate  (mg/L)	R	No. of Daphnids per replicate at start 0 h	Signs of toxicity and Immobilisation of Daphnids Observed at
				24 h		48 h
				Toxic signs	I	Toxic signs	I
G1	0.0	R1	5	N(5)	0	N(5)	0
		R2	5	N(5)	0	N(5)	0
		R3	5	N(5)	0	N(5)	0
		R4	5	N(5)	0	N(5)	0
G2	0.010	R1	5	N(5)	0	N(5)	0
		R2	5	N(5)	0	N(5)	0
		R3	5	N(5)	0	N(5)	0
		R4	5	N(5)	0	N(5)	0
G3	0.021	R1	5	N(4), B(1)	0	N(3), B(1)	1
		R2	5	N(4), L(1)	0	N(3), B(1)	1
		R3	5	N(3), L(2)	0	N(3), L(2)	0
		R4	5	N(3), L(1), B(1)	0	N(2), L(1), B(2)	0
G4	0.047	R1	5	N(2), L(1), B(1)	1	N(2), L(1)	1
		R2	5	N(2), B(1)	2	N(1), L(1), B(1)	0
		R3	5	N(2), B(3)	0	N(2), B(2)	1
		R4	5	N(3), L(1)	1	N(2), L(1)	1
G5	0.103	R1	5	N(1), B(2)	2	B(2)	1
		R2	5	N(1), L(1), B(2)	1	L(1), B(1)	2
		R3	5	N(1), L(2), B(1)	1	L(1), B(1)	2
		R4	5	N(1), L(2)	2	L(2)	1
G6	0.227	R1	5	L(1), B(1)	3	L(1)	1
		R2	5	L(2), B(1)	2	B(1)	2
		R3	5	L(2), B(2)	1	B(1)	3
		R4	5	L(1), B(2)	2	B(1)	2
G7	0.500	R1	5	L(1), B(1)	3	-	2
		R2	5	L(1)	4	-	1
		R3	5	L(2), B(1)	2	-	3
		R4	5	L(1), B(1)	3	-	2

R: Replicate; h: hour; I: Immobilisation; N: Normal; B: Localized on Bottom; L: Lethargy.
Alphabet outside and values inside the parentheses represent the clinical symptom code and Number of Daphnids exhibiting that particular symptom, respectively; -: Not applicable.       

Table 2: TOC analysis during main study

Group	Nominal Loading rates  (mg/L)	TOC During Exposure (mg)
		Day 0 (Fresh)	Day 1 (Aged)	Day 1 (Fresh)	Day 2 (Aged)
G1	0.0	-	-	-	-
G2	0.010	0.1128	0.1103	0.1205	0.1186
G3	0.021	0.1619	0.1423	0.1689	0.1681
G4	0.047	0.2525	0.2486	0.2612	0.2601
G5	0.103	0.3387	0.3296	0.3412	0.3386
G6	0.227	0.3747	0.372	0.3795	0.3743
G7	0.500	0.4005	0.3985	0.4028	0.4008

-: Not Applicable.

 

 

Validity criteria fulfilled:

yes

Conclusions:

In the acute immobilisation test according to OECD guideline 202 the 48 hours acute median Effective Loading Rate (EL50) of Humectol LYS MOD on Daphnia magna is 0.0815 mg/L, the 95% confidence interval with lower limit of 0.0696 mg/L and upper limit of 0.0954 mg/L. No Observed Effect Loading Rate (NOELR) and Lowest Observed Effect Loading Rate (LOELR) over the 48 hours exposure period are 0.021 and 0.047 mg/L of Humectol LYS MOD respectively.

Executive summary:

The test item Humectol LYS MOD was evaluated for acute toxicity on Daphnia magna according to OECD Guideline No. 202. The acute toxicity study of Humectol LYS MOD on Daphnia magna was performed to determine the 48 hours Effective concentration (EL50) and to determine Lowest Observed Effect Nominal Loading Rate (LOELR) and No Observed Effect Nominal Loading Rate (NOELR). Daphnids were exposed for the test item by aqueous exposure (treatment) under semi-static condition. Daphnids were observed at 24 and 48 hours for signs of toxicity and immobilization.
Based on the results of a dose range finding study, main study was conducted at the nominal loading rates of 0.010, 0.021, 0.047, 0.103, 0.227 and 0.500 mg/L of Humectol LYS MOD of test medium along with control. No clinical signs of toxicity were observed in control and at the nominal loading rate of 0.010 mg/L during the 48 hours observation period. Clinical signs of daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 0.021, 0.047, 0.103 and 0.227 mg/L during the 48 hours observation period. Clinical signs of daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 0.500 mg/L during the 24 hours observation period. The immobilization percentages of 0, 10, 35, 60, 80 and 100% were observed at the nominal loading rates of 0.010, 0.021, 0.047, 0.103, 0.227 and 0.500 mg/L respectively during the 48 hours observation period.
Based on the above results of the experiment and under experimental conditions employed, it is concluded that the 48 hour acute median effective concentration (EL50) of Humectol LYS MOD on Daphnia magna is 0.0815 mg/L; the 95 % confidence interval with lower limit of 0.0696 mg/L and upper limit of 0.0954 mg/L. NOELR and LOELR over the 48 hours exposure period are 0.021 and 0.047 mg/L of Humectol LYS MOD, respectively.

Description of key information

In the acute immobilisation test according to OECD guideline 202 the 48 hours acute median Effective Loading Rate (EL50) of Humectol LYS MOD on Daphnia magna is 0.0815 mg/L, the 95% confidence interval with lower limit of 0.0696 mg/L and upper limit of 0.0954 mg/L. No Observed Effect Loading Rate (NOELR) and Lowest Observed Effect Loading Rate (LOELR) over the 48 hours exposure period are 0.021 and 0.047 mg/L of Humectol LYS MOD respectively.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.082 mg/L

Additional information

The test item Humectol LYS MOD was evaluated for acute toxicity on Daphnia magna according to OECD Guideline No. 202. The acute toxicity study of Humectol LYS MOD on Daphnia magna was performed to determine the 48 hours Effective concentration (EL50) and to determine Lowest Observed Effect Nominal Loading Rate (LOELR) and No Observed Effect Nominal Loading Rate (NOELR). Daphnids were exposed for the test item by aqueous exposure (treatment) under semi-static condition. Daphnids were observed at 24 and 48 hours for signs of toxicity and immobilization.
Based on the results of a dose range finding study, main study was conducted at the nominal loading rates of 0.010, 0.021, 0.047, 0.103, 0.227 and 0.500 mg/L of Humectol LYS MOD of test medium along with control. No clinical signs of toxicity were observed in control and at the nominal loading rate of 0.010 mg/L during the 48 hours observation period. Clinical signs of daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 0.021, 0.047, 0.103 and 0.227 mg/L during the 48 hours observation period. Clinical signs of daphnids localized on bottom of container and lethargy was observed at the nominal loading rates of 0.500 mg/L during the 24 hours observation period. The immobilization percentages of 0, 10, 35, 60, 80 and 100% were observed at the nominal loading rates of 0.010, 0.021, 0.047, 0.103, 0.227 and 0.500 mg/L respectively during the 48 hours observation period.
Based on the above results of the experiment and under experimental conditions employed, it is concluded that the 48 hour acute median effective concentration (EL50) of Humectol LYS MOD on Daphnia magna is 0.0815 mg/L; the 95 % confidence interval with lower limit of 0.0696 mg/L and upper limit of 0.0954 mg/L. NOELR and LOELR over the 48 hours exposure period are 0.021 and 0.047 mg/L of Humectol LYS MOD, respectively.