Alcohols, C12-14, ethoxylated

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 Feb - 26 Mar 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations
- Sampling method: samples were taken at test start, after 24 h fresh and aged and after 48 and 72 h from fresh solutions. For nominal test item concentration 3.41 and 7.50 mg/L analytical samples from t = 0 h fresh and t = 24 h aged were analysed as at this highest test concentration all fish were dead after 4-6 h.
- Sample storage conditions before analysis:The samples were mixed with acetonitrile and stored deep-frozen (≤ - 18 °C) until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared by adding the necessary amount of test item to a volumetric flask. Test medium was added and the solution was stirred for 1 h at 1000 rpm. The solution was observed to be turbid. Test item concentrations were prepared by dilution. The two highest test concentrations were observed to be turbid, all other solutions were observed to be clear and transparent. All test solutions were foamy.

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Source: bred at testing facility
- Weight at study initiation (mean and range, SD): average: 0.3868  0.1336 g

ACCLIMATION
- Acclimation conditions (same as test or not): same
- Type and amount of food during acclimation: commercial fish food (inicio plus, BioMar SAS)
- Feeding frequency during acclimation: once per day, last feeding was performed two days prior exposure start
- Health during acclimation (any mortality observed): no

FEEDING DURING TEST (as applicable)
none

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

179 mg CaCO3/L

Test temperature:

21.5 – 22.7 °C

pH:

7.46 – 7.77

Dissolved oxygen:

63- 119%

Conductivity:

432 µS/cm

Nominal and measured concentrations:

Nominal test substance concentrations: 0.320, 0.704, 1.55, 3.41, 7.50 mg/L
Time weighted mean measured concentrations: 0.0898, 0.158, 0.304, 0.640, 5.60 mg/L (based on 2 representative contituents)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria (18 L nominal volume)
- Type (delete if not applicable): covered with lids
- Fill volume: 15 L
- Aeration: none
- Renewal rate of test solution (frequency): every 24 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.0785 g fish/L (control)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated drinking water and deionised water
- Total organic carbon: ≤ 2.0 mg/L (non-GLP)
- Culture medium different from test medium: no
- Intervals of water quality measurement: 0 hours from fresh test solutions and after 24, 48, 72 and 96 hours from fresh and aged test solutions. Oxygen saturation was monitored after 2 h, 4 h and 6 h.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light/ 8 h darkness
- Light intensity: 830 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and toxicity symptoms after 0 h, 4-6 h, 1 day (24 h), 2 days (twice per day), 3 days (twice per day) and 4 days (96 h) of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range finding study
- Test concentrations: 0.01, 0.1, 1.0, 10.0 mg/L
- Results used to determine the conditions for the definitive study: 100% mortality at 10 mg/L

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.423 mg/L

95% CI:

>= 0.327 - <= 0.594

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on two representative constituents

Remarks:

C12EO2, C12EO3

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.304 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on two representative constituents

Remarks:

C12EO2, C12EO3

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: No sublethal effects were observed in the control and at test item concentrations up to and including 0.704 mg/L (nominal). After 4 – 6 hours, two fish showed reduced activity at test item concentration 1.55 mg/L (nominal) and one fish were in lateral position at the bottom of the glass aquaria at test item concentration 3.41 mg/L.
- Mortality of control: no
- Other adverse effects control: no
The test item solutions in the glass aquaria were clear and transparent, except for nominal test item concentrations of 3.41 and 7.50 mg/L, which were foamy and turbid.

Reported statistics and error estimates:

The NOEC and LOEC were determined by using a multiple comparison method (step down Cochran-Armitage test procedure). The LC50-value after 4 - 6 h was determined by logit analysis using linear maximum likelihood regression and the 24 h, 48 h, 72 h and the 96 h LC50-values were determined by probit analysis using linear maximum likelihood regression.

Sublethal observations / clinical signs:

Table 1: Validity criteria for OECD 203 (2019)

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	No mortality observed	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	≥73%	yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80% of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20 per cent, results should be based on the measured concentration.	analytical monitoring performed, results are based on TWA	yes

 

Table 2: [% ] mortality of fish

Test item nominal conc.[mg/L]	Control	0.320	0.704	1.55	3.41	7.50
Time [h]	Mortality [%]
4 – 6	0	0	0	0	86	100
Day 1 (24)	0	0	0	0	100	100
Day 2 (48)	0	0	0	0	100	100
Day 2 (51 - 54)	0	0	0	0	100	100
Day 3 (72)	0	0	0	0	100	100
Day 3 (75 - 78)	0	0	0	0	100	100
Day 4 (96)	0	0	0	0	100	100

Seven fish were used for each replicate in the test

Table 3: Observation of clinical signs of fish during testT

est item nominal conc.[mg/L]	Control					0.320					0.704
Time [h]	0	°	*	#	+	0	°	*	#	+	0	°	*	#	+
4 – 6	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Day 1 (24)	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Day 2 (48)	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Day 2 (51 - 54)	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Day 3 (72)	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Day 3 (75 - 78)	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Day 4 (96)	7	0	0	0	0	7	0	0	0	0	7	0	0	0	0
Test item nominal conc.[mg/L]	1.55					3.41					7.50
Time [h]	0	°	*	#	+	0	°	*	#	+	0	°	*	#	+
4 – 6	5	2	0	0	0	0	0	0	1	6	0	0	0	0	7
Day 1 (24)	7	0	0	0	0	0	0	0	0	7	0	0	0	0	7
Day 2 (48)	7	0	0	0	0	0	0	0	0	7	0	0	0	0	7
Day 2 (51 - 54)	7	0	0	0	0	0	0	0	0	7	0	0	0	0	7
Day 3 (72)	7	0	0	0	0	0	0	0	0	7	0	0	0	0	7
Day 3 (75 - 78)	7	0	0	0	0	0	0	0	0	7	0	0	0	0	7
Day 4 (96)	7	0	0	0	0	0	0	0	0	7	0	0	0	0	7

0 = no clinical signs

° = unusual behaviour (reduced activity and/or orientation to bottom or surface of the test vessels, dark pigmentation, dark eyes)

* = difficulties with maintenance of equilibrium

# = fish upside down with loss of equilibrium, showing only movement of gills as a sign of life

+ = no sign of life

Validity criteria fulfilled:

yes

Remarks:

Please see table 1 at "any other information on results incl. tables"

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

07 - 11 NOV 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1984

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations
- Sampling method: samples were taken from fresh solutions and from aged solutions at 0, 24, 48 and 72 h

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1 g of test substance/L was diluted in demineralized water and stirred over 18 h. The test solution was filtered.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: West Aquarium, Bad Lauterberg, Germany
- Length at study initiation (length definition, mean, range and SD): 3 cm +/- 0.5 cm
- Feeding during test: no feeding


ACCLIMATION
- Acclimation period: 14 days quarantine after treatment with malachit-green
- Acclimation conditions (same as test or not): dechlorinated drinking water in 200 L stone ware aquaria or 300 L glas aquaria, 20 °C +/- 2 °C, flow-
through conditions
- Type and amount of food: TetraMin approx. 1% of body weight
- Feeding frequency: daily
- Health during acclimation (any mortality observed): animals were healthy at test start, mortality <= 5% prior test start

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

10.8 ° dH (2nd test), 10.5 ° dH (3rd test)

Test temperature:

20 °C

pH:

7.8 - 8.3 (2nd test)
7.8 - 8.4 (3rd test)

Dissolved oxygen:

80 - 101% (2nd test)
98 - 106% (3rd test)

Nominal and measured concentrations:

nominal: 0.12, 0.25, 0.50, 1.0, 2.0 mg/L (2nd test) and 3.6 and 6.0 mg /L (3rd test)
mean measured (geometric mean): 0.12, 0.12, 0.13, 0.46, 0.66 mg/L (2nd test) and 2.4 and 4.4 mg/L (3rd test)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria (20 L)
- Fill volume: 10 L
- Aeration: continuous
- Renewal rate of test solution (frequency): daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.25 g/fish (2. test), 0.20 g/fish (3. test)


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking water (Gelsenwasser AG)
- Intervals of water quality measurement: daily


OTHER TEST CONDITIONS
- Photoperiod: 16 light / 8 h darkness

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.2 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.66 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: no

Sublethal observations / clinical signs:

Table 1: Mortalities

Mean measured concentration [mg/L]	Test period (h)
	24 h	48 h	72 h	96 h
	Mortality [%]	Mortality [%]	Mortality [%]	Mortality [%]
2nd test
Control	0	0	0	0
0.12	0	0	0	0
0.12	0	0	0	0
0.13	0	0	0	0
0.46	0	0	0	0
0.56	0	0	0	0
3rd test
Control	0	0	0	0
2.4	100	100	100	100
4.4	100	100	100	100

Table 2: Validity criteria for OECD 203 (2019)

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	80 - 101% (2nd test) 98 - 106% (3rd test)	yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80% of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20 per cent, results should be based on the measured concentration.	results based on measured concentration

 

Validity criteria fulfilled:

yes

Remarks:

Please refer to table 2 at "any other information on results incl. tables"

Conclusions:

The 96-hour LC50 of Alcohols, C12-14, ethoxylated (2 EO) to Brachydanio rerio was investigated in accordance to EU Guideline 92/69 EWG Part C 1. The LC50 was determined to be 1.2 mg/l.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

01 - 05 JUL 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1981

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution of the test substance was prepared at a concentration of 2 mg/L in the water used for the test. 10 ml of the test solution was introduced into the 10-litre tanks used in the test and kept stirring for 1 hour before the fish were introduced.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Acquariomania, Milano, Italy
- Length at study initiation (length definition, mean, range and SD): 3 cm ± 0.5 cm
- Weight at study initiation (mean and range, SD): 400 mg ± 100 mg


ACCLIMATION
- Acclimation period: 15 days
- Type and amount of food during acclimation: Tetreamin fishfood (Tetrawerke, Germany)
- Feeding frequency during acclimation: daily unitl 24 before test start
- Health during acclimation (any mortality observed): no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

180 mg/CaCO3/L

Test temperature:

22 ± 2 °C

pH:

7 ± 0.1

Dissolved oxygen:

6.6 mg/L

Nominal and measured concentrations:

Nominal test substance concentration: 2 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria (10 L nominal volume)
- Fill volume: 10 L
- Aeration: continous
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 500 mg/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filtered tap water

OTHER TEST CONDITIONS
- Photoperiod: 12 h light/12 h darkness
- Light intensity: fluorescent light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and symptoms of intoxication after 2-4, 8 h and subsequently every 24 h

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: no
- Other biological observations: no symptoms observed.

Sublethal observations / clinical signs:

Table 1: Results

Exposure time [h]	Symptoms	Mortality [%]
2	none	0
4	none	0
8	none	0
24	none	0
48	none	0
72	none	0
96	none	0

Validity criteria fulfilled:

not specified

Description of key information

LC50 (96 h): 0.423 mg/L (TWA; OECD 203).

Additional information

The acute toxicity of Alcohols, C12-14, ethoxylated to fish was investigated in a semi-static test design following OECD guideline 203. The fish (Pimephales promelas) were exposed to nominal test substance concentration series of 0.320, 0.704, 1.55, 3.41, and 7.50 mg/L with a daily renewal of test solutions. the test concentrations were analytically verified by HPLC-MS/MS.The time weighted mean measured concentrations, based on 2 representative constituents of the UVCB substance were 0.0898, 0.158, 0.304, 0.640, and 5.60 mg/L. The determined LC50(96 h) was 0.423 mg/L (TWA).

Two supporting studies conducted with Alcohols, C12-14, ethoxylated are available. Both studies were conducted according to OECD 203 with Danio rerio as test species. One of the studies used a semi-static study test design with a daily renewal of the test solution. The determined LC50 (96 h) was 1.2 mg(L (meas. geom. mean). the second supporting study was conducted as a static limit test with a nominal concentration of 2 mg/L. The determined LC50 (96 h) was > 2.0 mg/L (nominal).