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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 March 1997 to 27 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 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples were taken from the 20 and 2 mg/mL methanol stock solutions after preparation. No analysis of test media was possible.

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 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):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: no data
- Source: (laboratory, culture collection) originally obtained via Shell Research, Sittingbourne UK
- Age at study initiation (mean and range, SD): < 24 h, collected from the third brood onwards from adults aged between 14 and 42 days, only cultures without ephippia were used.
- Method of breeding: cultured in 600-mL glass beakers containing 500 mL of reconstituted water. Each vessel and its contents is considered a 'culture'. New cultures were started with D. magna less than 24 hours old, at a density of about 20 per litre.
- Feeding during test
- Food type: concentrated suspension of Chlorella vulgaris (5 x 10^5 cells/mL, obtained from 5-L culture grown under semi-axenic conditions, concentrated by filtration and resuspended in reconstituted freshwater) and yeast (Allinson's Dried Bakers Yeast)
- Amount: 0.2 mL (50 mg/L suspension) per day
- Frequency: daily

ACCLIMATION
- Acclimation period: 1 h

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

not applicable

Hardness:

see Table 1

Test temperature:

see Table 1

pH:

see Table 1

Dissolved oxygen:

see Table 1

Salinity:

not applicable

Nominal and measured concentrations:

nominal concentration of methanol stock solution: 2 mg/mL and 20 mg/mL
nominal concentration of test medium: 0.2 mg/L
mean measured concentrations of methanol stock solutions (0 h and 24 h): 2.059 mg/mL and 18.426 mg/mL (103 % and 92 %, 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: dishes
- Material, size, headspace, fill volume: glass cristallising
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): transfer of D. magna to new vessel containing freshly prepared media
- Renewal rate of test solution (frequency/flow rate): not applicable
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4 containing dilution water only
- No. of vessels per vehicle control (replicates): 4 containing methanol only at the same concentration as the test media,

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconsituted freshwater prepared by mixing treated mains water and a proprietary mineral water in the ratio 1:1
* treated mains dilution water: laboratory main supply. The water is pumped to the laboratory via a Purite AC9 activated carbon filter and a Purite ROPF 20 particulate filter.
* proprietary mineral water: Ashbourne, Still Natural Water (Nestle UK Ltd.)
- Water parameters measured: see table 1
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light : 8 hours dark
- Light intensity: fluorescent lighting

MONITORING:
- pH, temperature and concentration of dissolved oxygen determined in each test medium at the start and end of each exposure period
- total hardness and alkalinity of control determined at the start and end of each exposure period

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- immobilisation: D. magna were considered to be immobile if, when the contents of the test vessel were briefly swirled, they did not swim during a 15 second period of observation.

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 immobility to D. magna after a 48 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:

EC50

Effect conc.:

> 0.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

no abnormalities observed

Results with reference substance (positive control):

not applicable

Results of the study are based on nominal concentrations because determination of test substance in test media was not possible.

The lowest concentration causing 100 % immobilisation was not determined and the highest concentration causing no immobilisation was 0.2 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

Reliable study (Klimisch 1) reporting EC50 > 0.2 mg/L for Daphnia magna. Test circumstances showed no deviations from the OECD test protocol.

Description of key information

One GLP study was presented.

The Covance study conducted a guideline study (OECD 202/EC C.2) by exposing Daphnia magna for 48h in a semi-static limit test to a nominal concentration of 0.2 mg/L of Wingstay L-HLS. The EC50 was >0.2 mg/L and the NOEC was 0.2 mg/L, based on nominal concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

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 92 -103% of nominal.

No immobilisation occurred and no other effects were observed. It can be concluded that the substance does not cause any adverse effects to Daphnia magna at concentrations which are higher than its water solubility.