Alcohols, C12-13, branched and linear, 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:

20 Nov 2020 - 01 Feb 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 10.0 and 5.0 mg/L analytical samples from t = 0 h fresh and t = 24 h aged were analysed as at these test concentrations all fish were dead after 4-6 h.
- Sample storage conditions before analysis:The samples were mixed with 10 mL 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 homogenised by shaking. The solution was observed to be turbid. Test item concentrations were prepared by dilution with test medium. The two highest test concentrations were observed to be turbid, all other solutions were observed to be clear and transparent.

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: 0.7440 g (10 fish)

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:

196 mg CaCO3/L

Test temperature:

21.9 – 22.9 °C

pH:

7.50 – 7.95

Dissolved oxygen:

≥ 90 % of air saturation

Conductivity:

420 µS/cm

Nominal and measured concentrations:

Nominal test item concentrations: 10.0, 5.00, 2.50, 1.25, 0.625 mg/L
Time weighted mean concentrations: 5.70, 2.57, 1.47, 0.646, 0.316 mg/L (based on 3 representative constituents)

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: 0.0245 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: 738 Lux (mean)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and toxicity symptoms after 0 h, 4 -6 h, 24 h, 48 h, 51 – 54 h, 72 h, 75 – 78 h and 96 h of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range finding study
- Test concentrations: 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:

EC50

Effect conc.:

1.19 mg/L

95% CI:

>= 0.886 - <= 1.45

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on three representative constituents

Remarks:

C12EO2, C12EO3,and C12EO6

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.646 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

other: based on three representative constituents

Remarks:

C12EO2, C12EO3,and C12EO6

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 1.25 mg/L (nominal). At 2.50 mg/L (nominal) four showed reduced activity after 4 - 6 h and two fish after 24 h and 51 - 54 h.
- Mortality of control: no
- Other adverse effects control: no
The test item solutions in the glass aquaria were clear and transparent.

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 and 24 h were determined following the trimmed Spearman-Karber procedure and the LC50-value after 48 h, 72 h and 96 h were determined by Weibull 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	≥ 90%	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	yes

Table 2: % mortality of fish

Test item nominal conc.[mg/L]	Control	0.625	1.25	2.50	5.00	10.0
Time [h]	Mortality [%]
4 – 6	0	0	0	0	100	100
Day 1 (24)	0	0	0	43	100	100
Day 2 (48)	0	0	0	71	100	100
Day 2 (51 - 54)	0	0	0	71	100	100
Day 3 (72)	0	0	0	86	100	100
Day 3 (75 - 78)	0	0	0	86	100	100
Day 4 (96)	0	0	0	86	100	100

Seven fish were used for each replicate in the test

Table 3: Observation of clinical signs of fish during test

Test item nominal conc.[mg/L]	Control					0.625					1.25
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]	2.50					5.00					10.0
Time [h]	0	°	*	#	+	0	°	*	#	+	0	°	*	#	+
4 – 6	3	4	0	0	0	0	0	0	0	7	0	0	0	0	7
Day 1 (24)	2	2	0	0	3	0	0	0	0	7	0	0	0	0	7
Day 2 (48)	2	0	0	0	5	0	0	0	0	7	0	0	0	0	7
Day 2 (51 - 54)	0	2	0	0	5	0	0	0	0	7	0	0	0	0	7
Day 3 (72)	1	0	0	0	6	0	0	0	0	7	0	0	0	0	7
Day 3 (75 - 78)	1	0	0	0	6	0	0	0	0	7	0	0	0	0	7
Day 4 (96)	1	0	0	0	6	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:

12 SEP - 16 SEP 1994

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)

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Source: Aqua Ivrea, taly
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): 2.0 ± 1 cm
- Weight at study initiation (mean and range, SD): 0.35 ± 0.15 g

ACCLIMATION
- Acclimation period: >=7 days

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

23 - 24 °C

pH:

7.11 - 7.69 (control)
7.06 - 7.69 (treatment)

Dissolved oxygen:

6.0 - 8.4 mg/L (control)
5.0 - 8.4 mg/L (treatment)

Nominal and measured concentrations:

Nominal test substance concentrations: 1, 1.8, 3.2, 5.6 and 10 mg/L

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water
- Intervals of water quality measurement: every 24 h

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and symptoms at 0, 24, 48, 72and 96 h

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.8 - 3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: Decreased excape reflex was recorded at 24 h in all the fish treated with 1 and 1.8 mg/L. The fish recovered at the 48 h observation.
- Mortality of control: no
- Other adverse effects control: no

Sublethal observations / clinical signs:

Nominal concentration [mg/L]	Test period (h)
	24 h	48 h	72 h	96 h
	Mortality [%]	Mortality [%]	Mortality [%]	Mortality [%]
Control	0	0	0	0
1.0	0	0	0	0
1.8	0	0	0	0
3.2	100	100	100	100
5.6	100	100	100	100
10	100	100	100	100

Description of key information

LC50 (96 h): 1.19 mg/L (TWA).

Additional information

The acute toxicity of Alcohols, C12-13, branched and linear, ethoxylated to fish was investigated in a GLP guideline study following OECD 203. Seven fish of the species Pimephales promelas were exposed to nominal test item concentrations of 0.0625, 1.25, 2.50, 5.00 and 10.0 mg/L under semi-static conditions. Test solutions were renewed every 24 hours.

The test concentrations were analytically monitored by HPLC-MS/MS. The time weighted mean measured concentrations, based on three representative constituents of the UVCB substance, were 0.316, 0.646, 1.47, 2.57, 5.70 mg/L. No mortalities were observed up to and including a measured concentration of 0.646 mg/L. The determined LC50 (96 h) was 1.19 mg/L (TWA).

 

In a supporting study, performed according to OECD 203 with Danio rerio as test organism, a LC50 (96 h) between 1.8 – 3.2 mg/L (nominal) was recorded.