Phenol, 4-methyl-, reaction products with dicyclopentadiene and isobutylene

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:

24 March 1997 to 28 March 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD and EU), according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples of the 20 and 2 mg/mL methanol stock solutions were taken for analysis each time they were prepared. It was not possible to measure the test substance in the test medium.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution was prepared by direct addition of test substance to methanol
- Controls: one containing methanol at the same concentration as the test medium and one containing dilution water only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): methanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.01 %
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no, during the test, all test media were clear, colourless solutions

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Strain: batch 56448
- Source: Fish Network Ltd. (Upper Hayne Farm, Blackborough, UK)
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, range and SD): mean of 10 test-fish: 5.6 cm (range 5.4 cm to 5.8 cm)
- Weight at study initiation (mean and range, SD): mean of 10 test-fish: 1.894 g (range 1.797 g to 1.991 g)
- Method of breeding:
* water in holding tanks was replaced continuously
* uneaten food and debris were cleaned from the tanks at least weekly
* fish were fed a proprietary food (Hatchery 800)
* fish mortalities were recorded daily: cumulative < 5 % over seven-day period preceding the test
- Feeding during test: no

ACCLIMATION
- Acclimation period: minimum 12 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food: proprietary food (Hatchery 800)
- Feeding frequency: no food in the 24 h proceeding the test
- Health during acclimation (any mortality observed): fish would not be used for testing if mortality exceeded 5% or if disease was apparent.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

not applicable

Hardness:

see Table 2

Test temperature:

see Table 2

pH:

see Table 2

Dissolved oxygen:

see Table 2

Salinity:

not applicable

Nominal and measured concentrations:

nominal concentration of methanol stock solutions: 2 mg/mL and 20 mg/mL
nominal concentration of test medium: 0.2 mg/L
measured concentrations of methanol stock solutions: 2.021 mg/mL and 18.524 mg/mL (101 % and 93 % of nominal concentration, respectively)
Concentrations of test substance could not be determined in test solutions, but only in stock solutions, immediately after preparation.

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria
- Type (delete if not applicable): closed, with perspex lids
- Material, size, headspace, fill volume: glass, 15-L, containing 14 L of test medium
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): at 24 h intervals
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory mains supply, water pumped to laboratory via a Purite AC9 activated carbon filter and a Purite ROPF 20 particulate filter and then heated to approximately 13 °C
- Chlorine: see Table 2
- Culture medium different from test medium: no
- Intervals of water quality measurement: see monitoring (below)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16:8, light:dark
- Light intensity: artificial, no other specifications reported

MONITORING
- Dissolved oxygen concentration and pH of media in each test vessel determined at start and completion of each 24 h exposure period in fresh and old media.
- Total hardness (mg/L as CaCO3) and residual chlorine concentration in the dilution water determined at start of each 24 h exposure period.
- Temperature (ambient, minimum and maximum) determined every 24 h in control test medium

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- toxic symptoms: at 1, 24, 48, 72 and 96 h fish exhibiting toxic symptoms were recorded. Symptoms classified into 4 categories:
1) no effects
2) mild toxic effects
3) severe toxic effects
4) dead

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: a combined limit and range-finding test was performed and resulted in no toxicity to O. mykiss after a 96 h exposure period. Therefore, no definitive test was performed and only results of limit test are reported.
- Range finding study: results not reported

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 0.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 0.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 0.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

no abnormalities observed

Results with reference substance (positive control):

not applicable

Sublethal observations / clinical signs:

Results of the study are based on nominal concentrations because determination of test substance in test media was not possible. No mortalities were observed in any fish exposed to Wingstay L-HLS throughout the duration of the test. The lowest concentration causing 100 % mortality was not determined and the highest concentration causing no mortality was 0.2 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

Reliable (Klimisch 1) study reporting a 96-H LC50 > 0.2 mg/L for Oncorhynchus mykiss.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

25 February 1991 to 8 April 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study according to DIN 38 412 (part 15) guidelines. Acceptable well-documented study report which meets the basic scientific principles. Exposure time was 48h instead of 96h.

Qualifier:

according to guideline

Guideline:

other: DIN 38 412, part 15

Principles of method if other than guideline:

The study was performed according to internal laboratory guidelines (SOP/91/F/07), which are based on DIN 38 412, part 15.
A preliminary test was performed with a saturated solution of the test substance and two dilutions. The test was repeated to confirm the results.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

At the start of the test, the concentration of test substance in the saturated solution was measured using DOC-analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1 g/L of test substance was dissolved by constant magnetic stirring during 24 hours in mains water (dechlorinated and filtered with an active carbon and gravel filter); at start of the test, the solution was treated with ultrasound during 2 minutes and subsequently filtered
- 2 dilutions were prepared: 1:10 and 1:100
- Controls: 1 control vessel, no details provided
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no data

Test organisms (species):

Leuciscus idus

Details on test organisms:

TEST ORGANISM
- Common name: Goldorfe
- Strain: no data
- Source: West Aquarium GmbH + Co. KG, Lauterberg, Germany
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, range and SD): 5.8 cm
- Weight at study initiation (mean and range, SD): 2.4 g
- Body index: 1.2
- Method of breeding: see Acclimation, no additional information reported
- Feeding during test: no

ACCLIMATION
- Acclimation period: minimum 14 days
- Acclimation conditions (same as test or not): no, in 250-L glass aquaria under flow through conditions at 20 °C
- Type and amount of food, feeding frequency: once a day, trout breeding feed (Fa. HEMO)
- Health during acclimation (any mortality observed): no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

not applicable

Hardness:

14 German degrees of general hardness (°dH)

Test temperature:

18.8-20.8 °C

pH:

8.05-8.44

Dissolved oxygen:

7.0-10.3 mg/L

Salinity:

not applicable

Nominal and measured concentrations:

Nominal concentration of 1 g/L and two dilutions, 1:10 (i.e. 100 mg/L) and 1:100 (i.e. 10 mg/L)
Concentrations of the test substance in the saturated solution were calculated through DOC determination. In the preliminary test the concentration was 2.8 mg/L (3.2 mg DOC/L) and in the main test 2.5 mg/L (2.9 mg DOC/L).

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria
- Type (delete if not applicable): not specified
- Material, size, headspace, fill volume: 2 L testmedium, glass aquaria, 12-L
- Aeration: no data
- 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: pro Fish testmedium, d.h. pro Konz.201
- Water parameters: no data
- Culture medium different from test medium: no data
- Intervals of water quality measurement: pH, temperature and dissolved oxygen were measured at start and after 24 and 48 hours

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16:8, light:dark
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- mortality, observed after 2-4 hours of beginning of test, and after 24 and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: factor 10
- Justification for using less concentrations than requested by guideline: no data
- Test concentrations: same for preliminary and final test
- Results used to determine the conditions for the definitive study: no mortality observed in any of the treatments, i.e. control, test substance at stock solution concentration and in dilutions of 1:10 and 1:100

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 2.8 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Remarks:

(preliminary test)

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 2.5 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Remarks:

(main test)

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

no abnormalities observed

Results with reference substance (positive control):

not applicable

Validity criteria fulfilled:

not specified

Conclusions:

Reliable study (Klimisch 2) reporting no mortality in L. idus after 48 h of exposure to nominal concentrations of Lowinox® CPL up to 1000 mg/L. Ideally, fish should have been exposed for 96 hours.

Description of key information

Two GLP studies are presented.

The Covance study conducted a guideline study (OECD 203/EC C.1) by exposing Oncorhynchus mykiss for 96h in a semi-static limit test to a nominal concentration of 0.2 mg/L of Wingstay L-HLS. The LC50 was >0.2 mg/L and the NOEC was 0.2 mg/L, based on nominal concentrations.

The IBR study was conducted according to DIN 38412 (part 15) and exposed Leuciscus idus for 48h in a static test to nominal concentrations of 1000, 100 and 10 mg/L of Lowinox CPL. The LC50 was >1000 mg/L and the NOEC was 1000 mg/L, based on nominal concentrations.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.2 mg/L

Additional information

Due to the low water solubility, the test substance in the Covance study was first dissolved in methanol (20 and 2 g/L stock solutions) and then used in the test medium to obtain a nominal concentration of 0.2 mg/L. No undissolved material was present. The concentrations in the stock solutions were measured and were 93 -101% of nominal.

In the IBR study, the test substance was dissolved in water (1 g/L stock solution), stirred for 24h, ultrasonicated for 2 min and subsequently filtered. Dilutions to nominal concentrations of 100 and 10 mg/L were then prepared. No details were provided on any undissolved material. Using DOC analysis and assumptions on the molecular weight of the substance, actual concentrations were estimated for the stock solution: 2.5 -2.8 mg/L.

No mortality occurred in both studies, and no other effects were observed. It can be concluded that the substance does not cause mortality in fish at concentrations which are higher than its water solubility.