Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and Ethanol, 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 4th - March 1st, 2013

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:

(1992)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

(2008)

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

Version / remarks:

(1996)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations sampled for analysis
Range-finding test: 100 mg/L
Final test: 0 (blank-control) and 100 mg/L
- Sampling
Frequency: at t=0 h and t=96 h
Volume: 2 ml
Singular samples were taken from all test solutions.
- Sample storage conditions before analysis: Samples were stored in a freezer until analysis (≤ -15°C). On the day of analysis, the samples were defrosted at room temperature. If necessary, the samples were diluted with ISO-medium to obtain concentrations within the calibration range.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The preparation of test solutions started with the highest test concentration. No special treatment other than a short magnetic stirring period was applied to fully dissolve the test substance in test medium. The lower test concentrations were prepared by subsequent dilutions in test medium.
- Controls: blank control (test medium without test substance or other additives)
- Evidence of undissolved material: The final test solutions were all clear and colourless.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Length at study initiation (mean and SD)
Range-finding test: 3.4 ± 0.2 cm
Final test: 2.4 ± 0.1 cm
- Weight at study initiation (mean and SD, ten fish of the batch used for the test, were weighed and measured prior to the start of the test.)
Range-finding test: 1.12 ± 0.21 g
Final test: 0.15 ± 0.04 g
- Feeding during test: No feeding from 24 hours prior to the test and during the total test period

ACCLIMATION
- Acclimation period: At least 12 days after delivery.
In the range-finding test fish were acclimatised for 5 days after receipt instead of at least 12 days as proposed by the protocol. The study intend was to assess the concentration range for the main study. This was not affected by the shorter acclimatisation period applied.
- Acclimation conditions (same as test or not): yes
- Type of food: Pelleted fish food (Cyprico Crumble Excellent (300-500 um), Coppens International bv, Helmond, The Netherlands)
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

180 mg CaCO3/L

Test temperature:

- Limit test
Start: 22.1°C
During the test period the temperature varied between 21.4 and 21.7°C

pH:

- Limit test
Start: 7.8
During the test period the pH varied between 7.5 and 7.8

Dissolved oxygen:

- Limit test (mg/L)
Start: 9.2-9.5
During the test temperature varied between 6.9 and 8.5

Nominal and measured concentrations:

- Nominal
Range-finding test: 0.1, 1, 10 and 100 mg/L
Limit test: 0 (blank-control) and 100 mg/L

- Measured: See the "Remarks on results including figures and tables" field for details.

Details on test conditions:

TEST SYSTEM
- Test vessel: All-glass, with a capacity of 6.5 L and filled with 5 L of test solution
- Aeration: No
- No. of organisms per vessel:
Range-finding test: 3
Limit test: 7
- No. of vessels per concentration (replicates):
Range-finding test: 1
Limit test: 1
- No. of vessels per control (replicates):
Range-finding test: 1
Limit test: 1
- Biomass loading rate:
Range-finding test: 0.21 g fish/litre
Limit test: 0.21 g fish/litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap-water purified by reverse osmosis
- Culture medium different from test medium: No
- Intervals of water quality measurement: Dissolved oxygen content, pH and temperature were measured daily in all vessels with surviving fish, beginning at the start of the test (day 0).


OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality and other effects at t = 3¼, 24, 48, 72 and 96 hours following the start of exposure. In addition, every afternoon from day 0 and every morning from day 1 to observe for any dead or severely distressed fish.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Limit test (100 mg/L)
- Range finding study
- Test concentrations: 0.1, 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol (PCP)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Range finding test: No mortality or clinical effects were observed at any of the test concentrations during the test period. The LC50 was above 100 mg/L.
- Limit test: No mortality or clinical effects were observed as was expected based on the results of the range-finding test.
- Acceptability of the test
1. No mortality was observed in the control at the end of the test.
2. Test conditions were maintained constant throughout the test.
3. The dissolved oxygen concentration has been at least 60% of the air saturation value throughout the test (>5 mg/L at 22°C).
4. The concentrations of the substance being tested have been at least 80% of the nominal concentration throughout the test.

Results with reference substance (positive control):

- Pentachlorophenol (PCP): The 96h-LC50 for carp was 0.17 mg/L (95% confidence interval between 0.15 and
0.27 mg/L).

Sublethal observations / clinical signs:

- Measured concentrations

Range finding test:

Concentrations of the test substance in test medium - range-finding test

Time of sampling [hours]	Date of sampling	Date of analysis^1	Concentration			Relative to nominal [%]	Relative to initial [%]
			Nominal [mg/L]	Analysed [mg/L]	Analysed corrected^2 [mg/L]
0	04 -Feb-2013	11 -Feb-2013	100	113	103	103
24	05 -Feb-2013	11 -Feb-2013	100	114	105	105	101
96	08 -Feb-2013	11 -Feb-2013	100	115	105	105	102

¹          Samples were stored in the freezer (≤ -15°C) until the day of analysis.

²          Corrected for the 109% mean recovery of the 100 mg/l procedural recovery samples analysed in this series.

Limit test:

Concentrations of the test substance in test medium - final test

Time of sampling [hours]	Date of sampling	Date of analysis^1	Concentration			Relative to nominal [%]	Relative to initial [%]
			Nominal [mg/L]	Analysed [mg/L]	Analysed corrected^2 [mg/L]
0	25 -Feb-2013	04 -Mar-2013	0	n.d.		n.a.
			100	114	102	102
96	01 -Mar-2013	04 -Mar-2013	0	n.d.		n.a.
			100	113	102	102	100

¹          Samples were stored in the freezer (≤ -15°C) until the day of analysis.

²          Corrected for the 111% mean recovery of the 100 mg/l procedural recovery samples analysed in this series.

n.d.    Not detected.

n.a.     Not applicable.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present test Accelerator (PT 25E or PT 25E/2) induced no visible or lethal effects in carp at an analytically confirmed concentration of 100 mg/L (NOEC). The 96h-LC50 exceeded 100 mg/L based on analytically confirmed exposure concentrations.

Executive summary:

In a 96-h acute toxicity study conducted according to OECD guideline 203, carp (Cyprinus carpio) were exposed to Accelerator (PT 25E or PT 25E/2) under static conditions at the following nominal concentration in a limit test: 0 (blank-control) and 100 mg/L.

Accelerator (PT 25E or PT 25E/2) induced no visible or lethal effects in carp at an analytically confirmed concentration of 100 mg/L. The 96h-LC50 exceeded 100 mg/L based on analytically confirmed exposure concentrations. The study is considered to be reliable without restrictions.

Description of key information

In a 96-h acute toxicity study conducted according to OECD guideline 203, carp (Cyprinus carpio) were

exposed to Accelerator (PT 25E or PT 25E/2). The 96h-LC50 exceeded 100 mg/L based on analytically

confirmed exposure concentrations. The study is considered to be reliable without restrictions.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

In a 96-h acute toxicity study conducted according to OECD guideline 203, carp (Cyprinus carpio) were exposed to Accelerator (PT 25E or PT 25E/2) under static conditions at the following nominal concentration in a limit test: 0 (blank-control) and 100 mg/L.

Accelerator (PT 25E or PT 25E/2) induced no visible or lethal effects in carp at an analytically confirmed concentration of 100 mg/L. The 96h-LC50 exceeded 100 mg/L based on analytically confirmed exposure concentrations. The study is considered to be reliable without restrictions.