3-(2,2-dimethyl-3-hydroxypropyl)toluene

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:

weight of evidence

Study period:

August 17th to August 21st 1987 (actual test)

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no analytical verification, but no loss of the test substance was seen in the algae test and as well in the suporting fish study. Thus, the results based on nominal values are reliable

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1984

GLP compliance:

yes

Analytical monitoring:

no

Remarks:

No loss of the test substance was seen in the algae test and as well in the suporting fish study.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The calculated amount of test sample was exactly weighed using an analytical balance and transferred quantitatively into a 2 L beaker. The suspension was stirred for 2 h using a magnetic stirrer. Immediately afterwards, the test sample was transferred quantitatively from the beaker into the test medium of one glass tank.

- Evidence of undissolved material (e.g. precipitate, surface film, etc.): For the test concentration of 1000 mg/L it could be observed that it led to a dispersion with visible particulate matter slowly settling down in the course of the test period.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra-fish
- Source: West Aquarium GmbH, Bad Lauterberg, Germany
- Age at study initiation (mean and range, SD): about 3 months
- Length at study initiation (length definition, mean, range and SD): average total length 2 +/- 1 cm

ACCLIMATION
- Acclimation period: 1987-07-24 to 1987-08-17
- Acclimation conditions (same as test or not): same as test, tank with 100L
- Type and amount of food during acclimation: Tetra Min Hauptfutter
- Feeding frequency during acclimation: twice daily, 24h before the test was started feeding was stopped
- Health during acclimation (any mortality observed): Good, no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20.4 - 21.4°C

pH:

7.9 - 8.1

Dissolved oxygen:

Oxygen concentration: 8.9 - 9.4

Nominal and measured concentrations:

Nominal concentrations: 1, 1.7, 2.9, 4.9, 8.3, 10, 100, 1000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass tanks
- Type (delete if not applicable): covered with glass plate assuring the aeration through pasteur pipettes as well as the free exchange with the surrounding atmosphere, but preventing effectively the fish to jump out of the test medium
- Material, size, headspace, fill volume: glass tanks: length 300 mm, width 200 mm and height 220 mm
- Aeration: yes
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: According to guideline
- Culture medium different from test medium: no
- Intervals of water quality measurement: at 0, 24, 48, 72 and 96 hours

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

4.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Remarks:

obtained graphically

Effect conc.:

9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none reported
- Observations on body length and weight: no
- Mortality of control: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: For the test concentration of 1000 mg/L it could be observed that it led to a dispersion with visible particulate matter slowly settling down in the course of the test period.
- Effect concentrations exceeding solubility of substance in test medium: yes, for 1000 mg/L (see above)

Sublethal observations / clinical signs:

Cumulative mortality at intervals during the test, cumulative percantage mortality in (%):

Concentration [mg/L]	Time after test start [h]
	0	1	24	48	72	96
1000	0*) (0)	10 (100)	10 (100)	10 (100)	10 (100)	10 (100)
100	0*) (0)	10 (100)	10 (100)	10 (100)	10 (100)	10 (100)
10	0*) (0)	0 (0)	0 (0)	10 (100)	10 (100)	10 (100)
8.3	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	1 (10)
4.9	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
2.9	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
1.7	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
1.0	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
Negative control	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)

*) Ten fish were taken at the beginning

Validity criteria fulfilled:

yes

Conclusions:

In the acute fish study with Danio rerio according to OECD 203 an LC50(96h) of 9 mg/L was determined.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Actual test: 25th January to 2nd February 1988

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:

1984

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples of water were taken from each test batch at the beginning, before the fishes were set into the batches, 24 h, 48 h and 96 h after test start to determine the concentration of the test sample in the test batches.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The calculated amount of test sample was exactly weighed using an analytical balance and transferred quantitatively into a 2 L beaker. The suspension (basic solution) was stirred for 2 h using a magnetic stirrer. Immediately afterwards, the test sample was transferred quantitatively from the beaker into the test medium of one glass tank.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra-fish
- Source: West Aquarium GmbH, Bad Lauterberg, Germany
- Age at study initiation (mean and range, SD): about 3 months
- Length at study initiation (length definition, mean, range and SD): average total length 2 +/- 1 cm

ACCLIMATION
- Acclimation period: 1987-07-24 to 1988-01-25
- Acclimation conditions (same as test or not): same as test, tank with 100L
- Type and amount of food during acclimation: Tetra Min Hauptfutter
- Feeding frequency during acclimation: twice daily, 24h before the test was started feeding was stopped
- Health during acclimation (any mortality observed): Good, no mortality observed

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20.3 - 22.3

pH:

7.6 - 7.7

Dissolved oxygen:

Oxygen concentration: 8.7 - 9.0

Nominal and measured concentrations:

Nominal concentrations: 0, 0.95, 1.7, 3.1, 5.6, 10, 18, 32 mg/L
Measured concentrations (96h): 0 (Control), 8.5 (nominal 5.6), 16 (nominal 10), 23 (nominal 18) mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass tanks
- Type (delete if not applicable): covered with glass plate assuring the aeration through pasteur pipettes as well as the free exchange with the surrounding atmosphere, but preventing effectively the fish to jump out of the test medium
- Material, size, headspace, fill volume: glass tanks: length 300 mm, width 200 mm and height 220 mm
- Aeration: yes
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: According to guideline
- Culture medium different from test medium: no
- Intervals of water quality measurement: at 0, 24, 48, 72 and 96 hours

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

5.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none reported
- Observations on body length and weight: no
- Mortality of control: none
- Measured concentrations: The measured concentrations after 96 hours were higher than the nominal concentrations / concentrations at t = 0h. Therefore the LC50 etc. were based on the nominal concentrations.

Sublethal observations / clinical signs:

Cumulative mortality at intervals during the test, cumulative percantage mortality in (%):

Nominal concentration [mg/L]	Time after test start [h]
	0	1	24	48	72	96
32	0*) (0)	8 (80)	10 (100)	10 (100)	10 (100)	10 (100)
18	0*) (0)	0 (0)	10 (100)	10 (100)	10 (100)	10 (100)
10	0*) (0)	0 (0)	0 (0)	1 (10)	1 (10)	1 (10)
5.6	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
3.1	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
1.7	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
0.95	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
Negative control	0*) (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)

*) Ten fish were taken at the beginning

Measured concentrations:

	Actual concentration [mg/L]
Nominal concentrations [mg/L]	0h	24h	48h	96h
18	22.9	22.8	- 1)	- 1)
10	10.7	8.7	16.0	16.0
5.6	6.0	4.8	7.5	8.5
0	0	0	0	-
Basic solution 1000	1030

1) LC100 was already obtained

Validity criteria fulfilled:

yes

Conclusions:

In the acute fish study with Danio rerio according to OECD 203 as flow-through test an LC50(96h) of 13 mg/L (nominal) was determined.

Description of key information

Geometric mean of two studies: LC50 = 10.8 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

10.8 mg/L

Additional information

Two experimental studies with the test substance studying the acute toxicity to Danio rerio according to OECD 203 are available. Both studies are in agreement with each other, showing similar results and were conducted under similar conditions.

The first study (1987, RL2) was done as a static test over 96h without analytical verification. As no loss of the test substance was seen in the algae test and as well in the second fish study, the results based on nominal values are reliable. Groups of 10 animals per concentration were exposed to the test substance at concentrations of 1, 1.7, 2.9, 4.9, 8.3, 10, 100 and 1000 mg/L in synthetic freshwater for 96 h. The highest tested concentration which did not cause mortality over 96h was 4.9 mg/L (=LC0), and the lowest test concentration leading to 100% mortality in the tested fish after 96 h of exposure was 10 mg/L (=LC100). The LC50 value, was graphically determined to be 9 mg/L.

The second study (1988, RL2) was done as a flow-through test over 96h including analytical verification. As the measurements of the test concentrations showed an increase of the concentrations over time, the nominal concentrations were used for determination of the LC50 etc (worst case). The test design was apart from the flow-through system analogue to the first study. Groups of 10 animals per concentration were exposed to the test substance at concentrations of 0.95, 1.7, 3.1, 5.6, 10, 18 and 32 mg/L in synthetic freshwater for 96 h. The highest tested concentration which did not cause mortality over 96h was 5.6 mg/L (=LC0), and the lowest test concentration leading to 100% mortality in the tested fish after 96 h of exposure was 18 mg/L (=LC100). The LC50 value, was graphically determined to be 13 mg/L.

As both studies were done with the same taxonomic group in the same lab and under similar conditions, both studies were used in a Weight of Evidence approach and the geometric mean is used as key value for the chemical safety assessment.