Reaction mass of methyl acetate and methanol

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

not specified

Remarks:

GLP-compliance not reported in the literature.

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

CULTURING
- Culturing: single clone Daphnia magna (clone K6) cultures were held in 1 L glass recipients filled with aerated and biologically filtered tap water, each containing 20 - 25 individuals.
- Conditions: the temperature in the temperature and light controlled chamber (Type WT150'/+5DU-WB, Weiss Technik, Liedekerke, Belgium) was maintained at 20 ± 1 °C with a photoperiod of 14 h light/10 h dark throughout culturing and exposure experiments.
- Water renewal: three times weekly
- Feeding: the daphnids were fed a mixture of P. subcapitata and Chlamydomonas reinhardtii in a 3/1 ratio (4 x 10E5 cells/ml)
- Age of daphnids at test initiation: < 24 h

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96

Hardness:

initial water hardness: 250 mg CaCO3

Test temperature:

20 ± 1 °C

pH:

initial pH: 7.8-8.2

Nominal and measured concentrations:

- Nominal concentrations (linear 1/2 dilution series): 1 - 32 g/L

Details on test conditions:

TEST SYSTEM
- Test vessels: glass recipients
- Test volume: 50 ml
- No of animals/replicate: 10
- No of replicates/concentration: 3
- Test medium: reconsituted water
- Test media renewal: every 24 h

TEST MEDIUM
- Medium: reconstituted water

TEST CONDITIONS
- Feeding: daphnids were fed the same algae mixture as during culturing
- Photoperiod: 14 h light/ 10 h dark

Key result

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

18 260 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

not specified

Conclusions:

 The 96-hour EC50 was determined to be 18260 mg/L (nominal).

Executive summary:

In a semi-static Daphnia sp. Acute Immobilisation Test according to OECD Guideline 202 young Daphnia magna (less than 24 hours old) were exposed for 96 hours to a linear ½ dilution series of 1 - 32 g Methanol/L in nominal. The test solutions were renewed every 24 hours. As a result, the 96-hour EC50 was determined to be 18260 mg/L (nominal).

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: DIN 38412 Teil 11

Principles of method if other than guideline:

German DIN guideline: "Bestimmung der Wirkung von Wasserinhaltsstoffen auf Kleinkrebse. Determination of the effects of substances in water to small crustacea

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Age at study initiation (mean and range, SD): 6 - 24 h
- Feeding during test: No feed was given during the test period.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

total hardness: 2.4 mmol/L
Calcium to magnesium ratio: 4:1
Sodium to potassium ratio: 10:1

Test temperature:

20°C

pH:

initial 8.0 ± 0.2

Dissolved oxygen:

≥ 2 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL beakers

- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: From the solution with the pollutant, dilution series were prepared using dilution water of the specified composition. The concentration steps of the test solution were selected so as to give 3-4 EC values in a range between EC0 and EC100, of which at least one value was below and one above EC50.

- acid capacity: 0.8 mmol/L

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

not specified

Conclusions:

The 48-hour EC50 was dtermined to be > 10000 mg/L.

Executive summary:

In a static test according to the German Guideline DIN 38412 with Daphnia magna the 48-hour EC50 was found to be > 10000 mg/L.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

not specified

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- additionally, tests with a second species, Daphnia pulex were conducted
- Source: Department of Biology, University of Turku, Finland
- Age at study initiation (mean and range, SD): < 24 h , neonates
- Method of breeding: cultured in M4 medium, 20 ± 2°C
- Food type: Scenedesmus algae every second or third day depending on the density of the culture.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Test temperature:

20 ± 2°C

Nominal and measured concentrations:

at least five concentrations; not specified

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 100 mL polypropylene vessels, with 50 mL liquid in each
- Aeration: no
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): at least 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Toxicity tests were performed in standard reference water (SRW; 2.4 mM NaHCO3 , 0.15 mM K2SO4 , 2.0 mM CaCI2 , 0.01 mM KH2PO4, pH 7.6) according to the OECD standard protocol with a few modifications.
- Photoperiod: 12:12-h light:dark

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

20 803 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The EC50 value for D. pulex were not obtained because of solubility problems (i.e., EC5O was not reached at the solubility limit of the chemical in the test solution, or the dose-response curves were irregular).

Reported statistics and error estimates:

The EC5O values were calculated using regression analysis after linearization of the dose-response curves by logarithmic transformation of the concentrations (statistical program; GraphPad 4, Intuitive Software for Science, San Diego, CA).

Validity criteria fulfilled:

not specified

Conclusions:

The 24-hour EC50 was determined to be 20803 mg/L.

Executive summary:

The acute toxicity of Methanol to Daphnia magna was assessed in a study according to OECD 202 under static conditions with at least five test item concentrations (not further specified) for a duration of 24 hours. The 24-hour EC50 was calculated to be 20803 mg/L.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

29.11.1994 - 22.12.1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Qualifier:

according to guideline

Guideline:

other: DIN 38 412, Part 11

GLP compliance:

yes

Remarks:

QAU statement available

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna Straus
- Strain: Klon 5
- Source: Daphnienzucht der Hoechst AG, Abteilung UWS
- Age at study initiation: > 6 h; < 23 h
- Method of breeding: Daphnids were held in 4 liter glass petri dishes. The medium was changed once per week and the animals were fed twice per day with algae (Scenedesmus subspicatus)
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20°C

pH:

7.7 - 7.8

Dissolved oxygen:

8.9 - 9.0

Nominal and measured concentrations:

Only nominal values are given

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: closed system, because of the volatility of the test item.
- Material, size, headspace, fill volume: 300 ml screw cap botteles made of glass. They were filled up to the brim with medium.
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2


TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: according to DIN 38412, part 30, and DIN 38412, part 11


OTHER TEST CONDITIONS
- Photoperiod: Test was performed in the dark


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility. Due to the fact that the test item was volatile and the test was therefore performed in closed vessels, effects were only determined after 48 hours.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: sqare root of 2

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

362 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1 026.7 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95 % confidence interval: 895.7 - 1435.4 mg/L

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

1 448.2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: immobility
- Mortality of control: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: 0.9 mg/l: 0 %; 1.9 mg/l: 75 %; 2.4 mg/l: 100 %
- EC50: > 0.9 mg/l < 1.9 mg/l

Reported statistics and error estimates:

A probit analysis was conducted for the estimation of the EC50 and the confidence intervals according to the method of Stephan (ASTM 634, American Society for Testing and Materials, 65-84, 1971)

Table 1: Effect of methyl acetate on immobilisation of Daphnia magna

Treatment [mg a.i./L] (nominal conc.)	No. of organisms at test begin	Observation period
		24 hours*		48 hours
		Cumul. no. of dead	% mortality	Cumul. no. of dead	% mortality
Control	70	—	—	0	0
128	20	—	—	0	0
181,02	20	—	—	0	0
256	20	—	—	0	0
362,04	20	—	—	0	0
512	20	—	—	1	5
724,08	20	—	—	1	5
1024	20	—	—	11	55
1448,16	20	—	—	20	100
2048	20	—	—	20	100

*not evaluated, because test was conducted in a closed system due to the volatility of the test item

 

Validity criteria fulfilled:

yes

Conclusions:

Based on the results, the EC50 of methyl acetate to Daphnia magna was calculated to be 1026.7 mg/L, based on nominal values.

Executive summary:

The 48–hr acute toxicity of methyl acetate to Daphnia magna was studied under static conditions.  Daphnids were exposed to control, and test chemical at nominal concentrations of 128, 181.02, 256, 362.04, 512, 724.08, 1024, 1448.16, and 2048 mg a.i./L} for 48 hr.  Immobilization was observed at test termination.  The 48– hour EC50 was 1026.7 mg a.i./L.  The 48– hr EC0 based on immobilization was 362.04 mg a.i./L.  This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Please refer to IUCLID section 13 for Read Across Justification.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

18 260 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Please refer to IUCLID section 13 for Read Across Justification.

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Reference 7

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Please refer to IUCLID section 13 for Read Across Justification.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

20 803

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: nominal/measured not specified

Reference 8

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Please refer to IUCLID section 13 for Read Across Justification.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1 026.7 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Description of key information

The EC50 of the registration substance was determined to be 1026.7 mg/L based on a read across to the structural analogue Methyl acetate (CAS 79-20-9).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1 026.7 mg/L

Additional information

No data on short-term toxicity to aquatic invertebrates are available for the registration substance itself, but for the read across source substances Methanol and Methyl acetate.

Methanol

In a semi-static Daphnia sp. Acute Immobilisation Test according to OECD Guideline 202 (Dom et al., 2012) young Daphnia magna (less than 24 hours old) were exposed for 96 hours to a linear 1/2 dilution series of 1 - 32 g Methanol/L in nominal. The test solutions were renewed every 24 hours. As a result, the 96-hour EC50 was determined to be 18260 mg/L (nominal).

In an additional study by Lilius et al. (1995) according to OECD 202 the acute toxicity of Methanol to Daphnia magna was assessed under static conditions with at least five test item concentrations (not further specified) for a duration of 24 hours. The 24-hour EC50 was calculated to be 20803 mg/L.

Further, in a static test according to the German Guideline DIN 38412 with Daphnia magna the 48-hour EC50 was found to be > 10000 mg/L (Kuehn et al., 1989).

Methyl acetate

The 48-hour acute toxicity of methyl acetate to Daphnia magna was studied under static conditions. Daphnids were exposed to control, and test chemical at nominal concentrations of 128, 181.02, 256, 362.04, 512, 724.08, 1024, 1448.16, and 2048 mg a.i./L for 48 hours.  Immobilization was observed at test termination. The 48-hour EC50 was 1026.7 mg a.i./L.  The 48-hour EC0 based on immobilization was 362.04 mg a.i./L (Stuhlfauth, 1994).  

Conclusion

Valid information from acute toxicity testing with Daphnia magna is available for the read across source substances Methanol and Methyl acetate. In studies according to OECD guideline 202 with Methanol (Dom et al., 2012; Kuehn, 1989) the EC50 was found to be >10000 g/L. Lilius et al. (1995) reported a 24-hour EC50 of 20803 mg/L. The 48-hour EC50 for Methyl acetate was determined to be 1026.7 mg a.i./L (Stuhlfauth, 1994). By applying a read across approach and under worst-case assumption the EC50 for the target substance is considered to be 1026.7 mg/L.