4-oxo-4-(p-tolyl)butyric acid adduct with 4-ethylmorpholine

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:

10.04.1995 - 14.04.1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

An amount of test material (2.00 g) was dissolved in 1 litre of dechlorinated tap water with the aid of ultrsonic disruption for approximately 45 minutes to give a 2.00 g/l stock solution which was then dispersed in a further 20 litres (final volume) of dechlorinated tap water to give the test concentration of 100 mg/l.
The concentration and stability of the test material in the test solutions were verified by chemical analysis at 0, 24 and 96 hours.

Vehicle:

no

Details on test solutions:

For the purpose of the definitive study the test material was prepared by direct dispersion in water.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The test was carried out using juvenile rainbow trout (Oncorhynchus mykiss). Fish were obtained from Donnington Fish Farm, Upper Swell, Gloucester, U. K. and maintained in-house since 21 February 1995. Fish were maintained in a glass fibre tank with a "single pass" water renewal system. Fish were acclimatised to test conditions from 3 April 1995 to 10 April 1995. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle.
The water temperature was controlled at 14 °C with a dissolved oxygen content of greater than or equal to 9.8 mg O2/l. These parameters were recorded daily. The stock fish were fed commercial trout pellets which was discontinued 48 hours prior to the start of the definitive study. There were zero mortalities in the 7 days prior to the start of the test and the fish had a mean standard length of 4.4 cm (s. d. = 0.2) and a mean weight of 0.42 g (s. d. = 0.03) at the end of the definitive study. Based on the mean weight value this gave a loading rate of 0.21 g bodyweight/litre.
The diet and diluent water are considered not to contain any contaminant that would affect the integrity and outcome of the study.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

4 d

Remarks on exposure duration:

After 3, 6, 24, 48, 72 and 96 hours any mortalities or adverse behavioural reactions to exposure were determined by visual inspection of the test fish

Hardness:

100 mg/l as CaCO3

Test temperature:

14 °C

pH:

7.4

Dissolved oxygen:

9.8 mg/l

Salinity:

Nitrate: 9.1 mg/l
Chloride: 35 mg/l
Sulphate: 59 mg/l
Fluoride: 314 mg/l
Magnesium: 9 mg/l
Sodium: 22 mg/l
Iron: 52 mg/l

Conductivity:

374

Nominal and measured concentrations:

With respect to benzenebutanoic acid, 4-methyl-γ-oxo:
Vessel 1:
0 hours: nominal: 100 mg/l; measured: 104 mg/l
24 hours: nominal: 100 mg/l; measured: 102 mg/l
96 hours: nominal: 100 mg/l; measured: 102 mg/l

Vessel 2:
0 hours: nominal: 100 mg/l; measured: 105 mg/l
24 hours: nominal: 100 mg/l; measured: 102 mg/l
96 hours: nominal: 100 mg/l; measured: 101 mg/l

With respect to 4-ethylmorpholine:
Vessel 1:
0 hours: nominal: 100 mg/l; measured: 97.2 mg/l
24 hours: nominal: 100 mg/l; measured: 87.4 mg/l
96 hours: nominal: 100 mg/l; measured: 89.6 mg/l

Vessel 2:
0 hours: nominal: 100 mg/l; measured: 94.1 mg/l
24 hours: nominal: 100 mg/l; measured: 88.4 mg/l
96 hours: nominal: 100 mg/l; measured: 94.6 mg/l

Details on test conditions:

Based on the results of the range-finding study a "Limit test" was conducted for the definitive study at a test concentration of 100 mg/l to confirm that at the maximum test concentration given in the OECD/EEC guidelines, no mortalities or adverse reactions to exposure were observed.
20 litre glass exposure vessels were used for each test concentration. At the start of the study 10 fish were placed in each vessel at random, in the prepared test solutions. The test vessels were then covered to reduce evaporation and maintained at 14°C with a photoperiod of 16 hours light and 8 hours darkness for a period of 96 hours. The test vessels were aerated via narrow bore glass tubes. The fish were not individually identified and received no food during exposure. A semi-static test regime was employed in the study involving a daily renewal of the test material remained near nominal and to prevent the build up of nitrogeneous waste products.
The water temperature, pH and dissolved oxygen concentrations were recorded daily throughout the study. The measurements at 0 hours represent those of the freshly prepared test preparations while the subsequent measurements at 24, 48, 72 and 96 hours represent those of the used or 24-hour old test preparations.
Water samples were taken from the control and each replicate test vessel at 0, 24, and 95 hours for individual quantitative analysis of each of the chemical components in the test solutions.

Reference substance (positive control):

no

Key result

Duration:

3 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

6 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 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.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There were no mortalities in 20 fish exposed to a test concentration of 100 mg/l for a period of 96 hours. The results of the definitive study showed the highest test concentration resulting in 0% mortality to be greater than or equal to 100 mg/l, the lowest concentration resulting in 100% mortality to be greater than 100 mg/l and the No Observed Effect Concentration (NOEC) to be greater than or equal to 100 mg/l. The No Observed Effect Concentration is based upon zero mortalities and the absence of any behavioural responses to exposure at this concentration.
It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/l.
There were no behavioural responses observed in 20 fish exposed to a test concentration of 100 mg/l for a period of 96 hours.
Temperature was maintained at 14 °C throughout the study, while there were no treatment related differences for oxygen concentration, a difference in pH was observed in the fresh test preparations at 0 hours between control (7.2) and the 100 mg/l test concentrations (6.9 - 7.0). Compared to these 0 hour values a slight increase in pH was observed for the control and the 100 mg/l 24-hour old test preparations (at 24, 48, 72 and 96 hours). However the difference between the control and the 100 mg/l test concentration remained approximately the same.
Analysis of the test preparations at 0, 24 and 96 hours was carried out separately for the chemical components benzenebutanoic acid, 4-methyl-γ-oxo and 4-ethylmorpholine. The analysis showed measured test concentrations of both chemical components to be near nominal on all sampling occasions and so it was considered justifiable to estimate LC50 values in terms of nominal test concentrations only.

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test material to freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-hour LC50 of greater than 100mg/l. Correspondingly the No Observed Effect Concentration was greater than or equal to 100 mg/l.

Executive summary:

The 96 hour LC50 for the test material to rainbow trout (Oncorhynchus mykiss), based on nominal test concentrations was greater than 100 mg/l and correspondingly the No Observed Effect Concentration was greater than or equal to 100 mg/l.

It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/l.

Individual analyis of each of the chemical components in the test solutions at 0, 24 and 96 hours showed the measured test concentrations to be near nominal and so the results are based on nominal test concentrations only.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

14.02.1995 - 18.02.1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 (incl. QA statement)

Analytical monitoring:

no

Details on sampling:

An amount of test material (2.00 g) was dissolved in 1 litre of dechlorinated tap water with the aid of ultrsonic disruption for approximately 45 minutes to give a 2.00 g/l stock solution which was then dispersed in a further 20 litres (final volume) of dechlorinated tap water to give the test concentration of 100 mg/l.
The concentration and stability of the test material in the test solutions were verified by chemical analysis at 0, 24 and 96 hours.

Vehicle:

no

Details on test solutions:

For the purpose of the definitive study the test material was prepared by direct dispersion in water.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The test was carried out using juvenile rainbow trout (Oncorhynchus mykiss). Fish were obtained from Donnington Fish Farm, Upper Swell, Gloucester, U. K. and maintained in-house since 21 February 1995. Fish were maintained in a glass fibre tank with a "single pass" water renewal system. Fish were acclimatised to test conditions from 3 April 1995 to 10 April 1995. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle.
The water temperature was controlled at 14 °C with a dissolved oxygen content of greater than or equal to 9.8 mg O2/l. These parameters were recorded daily. The stock fish were fed commercial trout pellets which was discontinued 48 hours prior to the start of the definitive study. There were zero mortalities in the 7 days prior to the start of the test and the fish had a mean standard length of 4.4 cm (s. d. = 0.2) and a mean weight of 0.42 g (s. d. = 0.03) at the end of the definitive study. Based on the mean weight value this gave a loading rate of 0.21 g bodyweight/litre.
The diet and diluent water are considered not to contain any contaminant that would affect the integrity and outcome of the study.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

4 d

Remarks on exposure duration:

After 24, 48, 72 and 96 hours any mortalities or adverse behavioural reactions to exposure were determined by visual inspection of the test fish.

Hardness:

100 mg/l as CaCO3

Test temperature:

14 °C

pH:

7.4

Dissolved oxygen:

9.8 mg/l

Salinity:

Nitrate: 9.1 mg/l
Chloride: 35 mg/l
Sulphate: 59 mg/l
Fluoride: 314 mg/l
Magnesium: 9 mg/l
Sodium: 22 mg/l
Iron: 52 mg/l

Conductivity:

374

Nominal and measured concentrations:

1, 10 and 100 mg/l

Details on test conditions:

For each test concentration 10 fish were added to each 20 litre test and control vessel and maintained at 14 °C with a photoperiod of 16 hours light and 8 hours darkness for a period of 96 hours under semi-static test conditions.
Each vessel was covered to reduce evaporation.

Reference substance (positive control):

no

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)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

There were no adverse reations to exposure (other than death) during the range-finding study.
The results showed no mortalities at the test concentrations of 1, 10 and 100 mg/l.
Based on this information, a single test concentration, in duplicate, of 100 mg/l was selected for the definitive study. This experimental design conforms to a "Limit test" to confirm that at the maximum test concentration given in the OECD/EEC Guidelines no mortalities or adverse reactions to exposure were observed.

Validity criteria fulfilled:

yes

Conclusions:

There were no adverse reations to exposure (other than death) during the range-finding study.
The results showed no mortalities at the test concentrations of 1, 10 and 100 mg/l.

Executive summary:

Based on this information, a single test concentration, in duplicate, of 100 mg/l was selected for the definitive study. This experimental design conforms to a "Limit test" to confirm that at the maximum test concentration given in the OECD/EEC Guidelines no mortalities or adverse reactions to exposure were observed.

Description of key information

The acute toxicity of the test material to freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-hour LC50 of greater than 100mg/l. Correspondingly the No Observed Effect Concentration was greater than or equal to 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L