Fatty acids, C16-18 and C18-unsatd., reaction products with triethanolamine, di-Me sulfate-quaternized

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13.09.2004 - 22.12.2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Range-Finding Test:
1 mg/L, 10 mg/L and control were analysed. Samples were taken at 0 h (initial value) from fresh test solution and after 24 h, after 48 h and at termination of the test after 96 hours from aged test solution.

Main Test:
2.56 mg/L, 4.10 mg/L and control were analysed. Samples were taken at 0 h (initial value) from fresh test solution, after 48 h from fresh and aged test solution and after 24 h, 72 h and at termination of the test after 96 hours from aged test solution. Analysis of 4.10 mg/L was terminated after 48 hours, since mortality was 100 % after 48 hours of exposure.

Vehicle:

no

Details on test solutions:

The substance is reported to be dispersable which does not mean it is soluble. No information about the physcal state in the test solutions (dispersed or diluted) are available in the test report.
- Method: no data given
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no data given

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Strain: Oncorhynchus mykiss (Walbaum)
- Length at study initiation: 4 - 6 cm
- Feeding during test: no

ACCLIMATION
- Acclimation period: > 12 d
- Acclimation conditions: 300 L containers with continuous renewal of water ( 5 - 10 % per day) and permanent aeration
- Feeding: each day with granular rearing food (size 0) with approximately 2 % of their body-weight. The food was provided by the fish supplier.
- Mortality: Following a 48-hour settling-in period, mortality was recorded. No mortality above 5 % was observed throughout the acclimatisation period

HOLDING WATER:
Water composed of dechlorinated drinking water and deionised water. The pH value of the aerated water was 6.5 - 8.5. The dissolved oxygen was above 60 % saturation and the total hardness was between 140 - 268 mg/L (as CaC03), corresponding to 7.8 - 15°dH.

HOLDING CONDITIONS:
- Light: 12 to 16 hours photoperiod daily
- Temperature: 15-17 °C
- Oxygen: > 60 %

Test type:

semi-static

Water media type:

freshwater

Total exposure duration:

96 h

Hardness:

12°dH (213.6 mg/L as CaCO3)

Test temperature:

15.6 - 17.8 °C

pH:

8.11 - 8.54

Dissolved oxygen:

> 60% of air saturation (approx. 6.0 mg O2/L)

Nominal and measured concentrations:

control, 1.0, 1.6, 2.56, 4.1, 6.55, 10.55 mg/L (nominal)
The mean measured concentration was determined to be 59.0% of nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: stainless steel tanks
- Material, size, headspace, fill volume: 26 L stainless steel tanks, filled with 24 L
- Aeration: continuous aeration of the test tanks with a membrane pump using a Pasteur pipette
- Renewal rate of test solution: 48-hour renewal of test medium
- No. of organisms per vessel: 10 per concentration
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: max. loading of 1 g fish/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: composed from dechlorinated drinking water and deionised water

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and sublethal effects (visible abnormalities as: loss of balance, swimming behaviour, respiratory function, pigmentation and all other events)


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0.01, 0.1, 1.0, 10, 100 mg/L
- The results indicated, that the LC50 (96 h) is between 1 mg/L and 10 mg/L and therefore the main test was conducted accordingly.

OBSERVATION INTERVALS:
- Fish were observed at 3, 6, 24, 48, 72 and 96 hours after initiation of the test.
- Measurements of temperature, pH-value and dissolved oxygen concentration were performed in 24 hour intervals from fresh and aged test solutions. Hardness of the untreated control was determined at the beginning of the test.

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.51 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.91 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

2.42 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

1.91 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

1.91 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

2 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 1.82-2.19 mg/l

Details on results:

Results based on nominal concentrations:
96h-LC50 = 3.24 mg/l**
72h-LC50 = 3.24 mg/l**
48h-LC50 = 3.24 mg/l**
24h-LC50 = 3.39 mg/l* (95% CL: 3.09 - 3.72)
96h NOEC = 2.56 mg/l ***

* calculated using Spearman-Karber estimator
** calculated using the geometric mean between the LC0 and LC100
*** not specified

Reported statistics and error estimates:

The LCx-values were determined using Spearman-Karber estimator or calculated as geometric mean between the LC0 and LC100, using the mortality data observed at different times with the nominal concentrations of the test item solution.

Sublethal observations / clinical signs:

Test results:

Table 1: % mortality of fish in the main test

	Nominal concentration of partially unsaturated TEA-Esterquat [mg/L]
	control	1.0	1.6	2.56*	4.10*	6.55*	10.5*
Time [h]	0	0	0	0	0	0	40
3	0	0	0	0	10	40	100
6	0	0	0	0	90	100	100
24	0	0	0	0	100	100	100
48	0	0	0	0	100	100	100
72	0	0	0	0	100	100	100
96	0	0	0	0	100	100	100

* foaming of the test solution was observed.

After 96 hours no mortality was observed at the nominal concentration of 2.56 mg/L (1.51 mg/L mean measured conc.) and below. 100 % mortality of fish was observed at 4.10 mg/L (2.42 mg/L mean measured conc.) and above after 96 hours.

Sublethal effects as difficulties with maintainance of balance and unusual behaviour (reduced activity and or orientation to bottom or surface of the test vessels) were observed within the first three hours in the highest test concentration.

No lethal or sublethal effects were observed in the control and the test concentrations up to 2.56 mg/L (NOEC) (1.51 mg/L mean measured conc.)


Results of analytical monitoring:

Table 2: Determined concentration of partially unsaturated TEA-EQ during the main test as % of nominal concentration

	nominal concentration of partially unsaturated TEA-Esterquat [mg/L]
Samples	0	2.56*	4.1*
Time [h]	actual concentration of partially unsaturated TEA-Esterquat [% of nominal]
0 fresh	-	104	92.2
24 aged	-	55.9	76.1
48 aged	-	20.3	27.9
48 fresh	-	10.2	**
72 aged	-	71.9	**
96 aged	-	10.2	**
Mean	-	-	**
		43.9	65.4

-  not calculated
* foaming of test item solution was observed
** no application and sampling since mortality was 100 % after 48 hours

The initial measured concentrations of partially unsaturated TEA-Esterquat were 104 and 92.2% of nominal at the analysed concentration levels. Since the measured concentrations of partially unsaturated TEA-Esterquat in the water samples fell graduately below 80 %, the toxicological endpoints were additionally evaluated using mean measured concentrations. The mean measured concentration was determined to be 59.0 % of nominal concentrations.

Validity criteria fulfilled:

yes

Conclusions:

According to the OECD and 92/69/EEC guidelines this study can be regarded
as valid, since
- the mortality in the control did not exceed 10 % at the end of the test.
- the dissolved oxygen concentration was at least 60 % of the air
saturation value throughout the test.

Executive summary:

In a 96-h acute toxicity study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test), rainbow trouts (Oncorhynchus mykiss) were exposed to partially unsaturated TEA-Esterquat (CAS No. 157905-74-3) at nominal concentrations of 0 (control), 1.0, 1.6, 2.56, 4.1, 6.55, 10.55 mg/l under semi static conditions. The mean measured concentration was determined to be 59 % of nominal concentrations. The 96-h LC50 was 1.91 mg/l (based on mean measured concentration). Sublethal effects as difficulties with maintainance of balance and unusual behaviour (reduced activity and or orientation to bottom or surface of the test vessels) were observed within the first three hours in the highest test concentration

This study is classified as reliable without restriction and satisfies the guideline requirements for an acute toxicity study with fish.

Results Synopsis
Test organism size (mean length): 4 - 6 cm
Test Type: Semi-static

96 h LC50=1.91 mg/l (mean measured concentration)

96h NOEC=1.51 mg/l
Endpoint(s) Effected: mortality, sublethal effects

Description of key information

96 h LC50 = 1.91 mg a.i/L, based on mean measured concentration (OECD TG 203, Oncorhynchus mykiss); GLP; RL1

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.91 mg/L

Additional information

In a 96-h acute toxicity study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test), rainbow trouts (Oncorhynchus mykiss) were exposed to partially unsaturated TEA-Esterquat (CAS No. 157905-74-3) at nominal concentrations of 0 (control), 1.0, 1.6, 2.56, 4.1, 6.55, 10.55 mg/L under semi-static conditions. The mean measured concentration was determined to be 59 % of nominal concentrations. The 96-h LC50 was 1.91 mg/L (based on mean measured concentration). Sublethal effects as difficulties with maintainance of balance and unusual behaviour (reduced activity and or orientation to bottom or surface of the test vessels) were observed within the first three hours in the highest test concentration  

 

In a further study performed according to OECD Guideline 203 (Fish, Acute Toxicity Test) under semi-static conditions the LC50 (96 h) of partially unsaturated TEA-Esterquat was determined to be 2.1 mg/L for Danio rerio. The effect concentration (96h-LC50 = 2.6 mg/L) was corrected because the analytical recovery (HPLC) of test substance was below 80% of nominal concentration.

Partially unsaturated TEA-Esterquat was classified as toxic to Danio rerio by the study authors. However, according to CLP (EU-GHS), Regulation (EC) No 1272/2008 the criteria for acute aquatic toxicity of fish (96h LC50) is </= 1 mg/L.

 

There are two further reliable studies available for partially unsaturated TEA-Esterquat performed according to OECD Guideline 203 (Fish, Acute Toxicity Test) under semi-static conditions and a study according to OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study) under flow-through conditions, but all performed without analytical control. The LC50 (96 h) effect concentrations were 7.48 mg/L (test mat., nominal, based on a.i. > 80 % the 96h LC50 was > 5.98 mg a.i./L) for Danio rerio and 11.23 mg a.i./L and 14.45 mg a.i./L for Oncorhynchus mykiss each, respectively. The nominal effect concentrations were corrected for the content of active ingredient.

One study is available for Leuciscus idus (Golden orfe). In this study the 24 to 96 h LC 50 of TEA-Esterquat partially unsaturated was 5.1 mg test mat./L (nominal).

In a flow through test according to OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study, 1984) the 96 h and 14 d LC50 of partially unsaturated TEA-Esterquat were 4.9 mg/L each (nominal) for Danio rerio.

 

The lowest 96 h LC50 in fish of 1.91 mg/L obtained in rainbow trouts (Oncorhynchus mykiss) will be used for chemical safety assessment.