Icosyl acrylate

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

from 2011-10-07 to 2011-11-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Version / remarks:

2010

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)

Version / remarks:

2008

Deviations:

no

Principles of method if other than guideline:

Modification of the report: The purity of the test substance was carried out at GKC Competence Center Analytics BASF SE, Ludwigshafen, Germany, no. 11L00419. The result was: 93.7 corr. area % as a sum of three main compounds. Reason for Amendment: At the point of finalization of the report to this study it was not possible to give a statement about the purity of the test substance because the characterization of the test substance in the anaIytical laboratory of GKC Competence Center Analytics BASF SE, Ludwigshafen, Germany was not complete.
No deviations from the test guidelines or other incidents occurred during the course of the reported test which may have influenced the results.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Details on sampling:

not applicable

Vehicle:

no

Details on test solutions:

Application of the test substance: Direct addition by initial weight into the test flasks. The test substance was weighed in the required amounts for the test concentrations directly to the test vessels.

The test substance was added in the required amounts according to the test concentrations directly to the test vessels with about 234 mL demineralized water. Aliquots of the reference substance stock solution were dosed to the test vessels and made up with demineralized water to a volume 234 mL. 16 mL synthetic medium were dosed to each test vessel with test substance and reference substance afterwards. To prepare the blank control assays 234 mL of demineralized water and 16 mL synthetic medium were mixed. The pH-values in all test vessels were checked. An adjustment was not necessary.

Test organisms (species):

activated sludge, domestic

Details on inoculum:

- Origin: Activated sludge from the municipal wastewater treatment plant of Mannheim, Germany was collected on 16 Nov 2011 from the aeration tank of the plant.
- Preparation of inoculum for exposure: The activated sludge suspension was sieved with a fine woven mesh (mesh size about 1 mm). This suspension was pre-aerated over night at room temperature. At the next day the sludge suspension was washed once with tap water and the suspension was adjusted to 3 g/L dry matter.
An aliquot of this suspension was added to the test vessels to obtain a sludge concentration of 1.5 g/L dry substance.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test temperature:

20.3 - 20.5 °C

Dissolved oxygen:

Oxygen concentration during aeration: > 2 mg/L
Oxygen concentration immediately before measurement: > 7 mg/L

Nominal and measured concentrations:

Test concentrations: 1000, 500, 250, 125, 62.5 mg/L
No measured concentrations were available.

Details on test conditions:

TEST SYSTEM
- Test vessel: Test vessels: Glas-beakers (nominal volume 1L)
- No. of vessels per test substance concentration (replicates): 2
- No. of vessels per control (replicates): 4
- No. of vessels per reference substance concentration (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic medium: 16 mUtest vessel of 100-fold concentrated OECD medium
- Incubation time: 3 hours
- Test temperature: 20.3 - 20.5 °C
- Test vessels: Glas-beakers (nominal volume 1L)
- Test volume: 500 mL
- Synthetic medium: 16 mL/test vessel of 100-fold concentrated OECD medium
- Oxygen concentration during aeration: > 2 mg/L
- Oxygen concentration immediately before measurement > 7 mg/L
- Duration of the measurement of oxygen consumption: 8 - 10 min
- Sludge concentration in the test vessel: 1.5 g/L.
- Test concentrations: 1000, 500, 250, 125, 62.5 mg/L as nominal concentration based on test substance without correction of purity or composition.
and 100, 10, 1 mg/L as nominal concentration based on reference substance.
- Reason for seleetion of test eoneentrations: According to the test guidelines, at least 5 concentrations in a geometric series with a separation factor of =< 3.2 should be used, preferably encompassing the range from 10 - 80% inhibition.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Duration:

3 h

Dose descriptor:

other: EC 20

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

other: EC80

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

EC10

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Details on results:

In conclusion the 3 hours effect concentrations for the test item in the Activated Sludge Respiration Inhibition test are:
- EC10 > 1000 mg/L
- EC20 > 1000 mg/L
- EC50 > 1000 mg/L
- EC80 > 1000 mg/L

Results with reference substance (positive control):

- The EC50 of 3,5-dichlorophenol was 5.3 mg/L. The EC50 of the reference substance 3,5-dichlorophenol was in the range of 2-25 mg/L in 3 hours therefore the results with reference substance are valid.

Reported statistics and error estimates:

The consumption rates were used for the determination of the ECx by the probit method based on Finney (1971) with the software TOXRAT Professional 2.10 (2001-2010).
The effect concentrations were given with an accuracy of 2 significant digits.

Reference 2

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

from 2012-05-03 to 2012-05-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Version / remarks:

2010

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Test concentrations: 1000, 500, 250, 125, 62.5 mg/L as nominal concentration based on test substance without correction of purity or composition.
100, 10, 1 mg/L as nominal concentration based on reference substance.
Test replicates: 4 replicates for the Control, 2 replicates for each test substance concentration, 2 replicates for each reference substance concentration.

Vehicle:

no

Details on test solutions:

- Application of the test substance: Direct addition by initial weight into the test flasks. The test substance was weighed in the required amounts for the test concentrations directly to the test vessels.
- Reference substance preparations: 1000.0 mg of the reference substance was added to about 800 mL of deionized water and stirred at room temperature until the reference substance was completely dissolved. The pH value of the stock solution was measured and adjusted to 7.2 with 1 M sodium hydroxide solution. Following this the stock solution was made up to 1L with deionized water. The stock solution appeared colorless-clear and no undissolved reference substance was visible.

Test organisms (species):

activated sludge, domestic

Details on inoculum:

- Source:Activated sludge from the municipal wastewater treatment plant of Mannheim, Germany, collected on 16 May 2012 from the aeration tank of the plant.
- Preparation of inoculum for exposure:The activated sludge suspension was sieved with a fine woven mesh (mesh size about 1 mm) and pre-aerated for about 2 days. During this time the sludge was fed daily with the synthetic sewage (50 mL synthetic sewage feed/L activated sludge). This synthetic sewage corresponds to the synthetic medium used for the measurement of the oxgen consumption.
- Initial biomass concentration: At the day of exposure the sludge suspension was washed once with drinking water and the suspension was adjusted to 3 g/L dry weight.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test temperature:

20.8 – 21.1°C

Dissolved oxygen:

Oxygen concentration during aeration: > 2 mg/L
Oxygen concentration immediately before measurement: > 7 mg/L
Duration of the measurement of oxygen consumption: 8 - 10 min

Nominal and measured concentrations:

- Test concentrations: 1000, 500, 250, 125, 62.5 mg/L as nominal concentration based on test substance without correction of purity or composition.
100, 10, 1 mg/L as nominal concentration based on reference substance.

Details on test conditions:

TEST SYSTEM
- Incubation time: 3 hours
- Test vessel: Glas-beakers (nominal volume 1L)
- Aeration: Yes
- Test volume: 500 mL

TEST MEDIUM / WATER PARAMETERS
- Synthetic medium: 16 mL/test vessel of 100-fold concentrated OECD medium
- Oxygen concentration during aeration: > 2 mg/L
- Oxygen concentration immediately before measurement: > 7 mg/L
- Duration of the measurement of oxygen consumption: 8 - 10 min
- Sludge concentration in the test vessel: 1.5 g/L

TEST CONCENTRATIONS
- Spacing factor for test concentrations (SF:2): 1000, 500, 250, 125, 62.5 mg/L as nominal concentration based on test substance without correction of purity or composition. 100, 10, 1 mg/L as nominal concentration based on reference substance.
- Justification for using less concentrations than requested by guideline: According to the test guidelines, at least 5 concentrations in a
geometric series with a separation factor of ≤ 3.2 should be used, preferably encompassing the range from 10 – 80% inhibition

Reference substance (positive control):

yes

Remarks:

Name of reference substance: 3,5 dichlorophenol; Reference substance No.: 02/0308-4; Batch-Identification: A0298018; CAS No.: 591-35-5 Expiry date: 28 Nov 2016.

Duration:

3 h

Dose descriptor:

EC10

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

other: EC20

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

other: EC80

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Details on results:

The results in this study were consistent with the validity criteria with one exception. The mean oxygen uptake in the blank controls was lower than 20 mg/g*h (16 mg/g*h). The reference substance shows an EC50 in the specified range (usual range of EC50 in the laboratory was 4.5 -11.9 mg/L in the year 2012). In the year 2012 the used activated sludge which was collected from the same waste water treatment plant showed a maximum oxygen consumption of 35.3 mg/g*h and minimum oxygen consumption of 11.8 mg/g*h. The study is classified as valid.

Results with reference substance (positive control):

The reference substance shows an EC50 in the specified range (usual range of EC50 in the laboratory was 4.5 -11.9 mg/L in the year 2012). In the year 2012 the used activated sludge which was collected from the same waste water treatment plant showed a maximum oxygen
consumption of 35.3 mg/g*h and minimum oxygen consumption of 11.8 mg/g*h. For this the study is classified as valid.
No deviations from the test guidelines or other incidents occurred during the course of the reported test which may have influenced the results.
In order to verify that the microorganisms are responding normally to toxic stress, tests with a reference substance are conducted according to OECD 209 guideline. The results from the reference substance test are compared to EC50 values published in OECD 209 guideline, which represent the typical response range.
According to the test guideline OECD 209 the EC50 values of the reference substance 3,5-dichlorophenol should be in the range of 2-25 mg/L after 3 hours of incubation.
The EC50 of the reference substance 3,5-dichlorophenol was in the range of 2-25 mg/L in 3 hours.
These results indicate that the microorganisms are responding normally to toxicant stress.

Reference 3

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

Meets National Standard Methods (DIN) with acceptable restrictions Read across was performed with Laurylacrylat 12/14 (mixture of laurylacrylate (CAS No.: 2156-97-0) and tetradecylacrylate (CAS No.: 21643-42-5) . Please refer to IUCLID section 13 for read across justification.

Qualifier:

according to guideline

Guideline:

other: German national standard, DIN 38412 Part 27, draft (ROBRA test)

Deviations:

no

GLP compliance:

no

Analytical monitoring:

no

Details on test solutions:

10000 mg test substance/L (nominal) was prepared by using 100 mg/L Tween 80 as a solubilizer and stirring for a period of 2 hours. The corresponding required test concentrations were prepared by taking appropriate volumes of the stirring stock solution. The emulsion appeared milky white and cloudy. Without stirring, dilution water and test substance separated after 5 min. Therefore aliquots were taken under constant stirring.

Test organisms (species):

Pseudomonas putida

Details on inoculum:

TEST ORGANISN
- Pseudomonas putida
- Strain: DSM 50026
- Age of bacterial culture: 2 hours

Water media type:

freshwater

Test temperature:

25 °C

pH:

7.2 - 7.3

Nominal and measured concentrations:

The test concentrations were: 100, 1000 and 10000 mg/L. Additionally, the highest test concentration (10000 mg/L) was tested in dilution water without glucose. A blank control (containing medium only) and a solvent control (containing 100 mg/L Tween 80) were tested in parallel.

Details on test conditions:

TEST SYSTEM
- Aeration: The test solution was aerated for 30 minutes at 25 °C.
- pH: 7.2 - 7.3
- Temperature: 25 °C
- Photometrical measurement at 436 nm

TEST CONCENTRATIONS. The test concentrations were: 100, 1000 and 10000 mg/L. Additionally, the highest test concentration (10000 mg/L) was tested in dilution water without glucose. A blank control (containing medium only) and a solvent control (containing 100 mg/L Tween 80) were tested in parallel.

Duration:

30 min

Dose descriptor:

EC10

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

30 min

Dose descriptor:

EC0

Effect conc.:

>= 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

30 min

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Details on results:

The EC50 value was determined to be greater than 10000 mg/L and also the EC10 was determined to be greater than 10000 mg/L.

Description of key information

The toxicity to microorganisms is assessed based on data available for 2-Propenoic acid, C12-14-alkyl esters (CAS 84238-60-8),2-Propenoic acid, C16-18-alkyl esters (CAS 90530-21-5), and 2-Propenoic acid, C18-22-alkyl esters (85085-17-2.

 

Table 1. Results on toxicity to aquatic microorganisms

Ester	Method	Species	EC10/ EC20/ EC50	Reference
2-Propenoic acid, C12-14-alkyl esters [84238-60-8]	DIN 38412, part 27	Pseudomonas putida	> 10000 mg/L	BASF SE 1988
2-Propenoic acid, C16-18-alkyl esters [90530-21-5]	OECD 209	activated sludge	> 1000 mg/L	BASF SE 2012
2-Propenoic acid, C18-22-alkyl esters [85085-17-2]	OECD 209	activated sludge	> 1000 mg/L	BASF SE 2011

 

Conclusion

The absence of a toxic potential to aquatic microorganisms was demonstrated byassays for2-Propenoic acid, C12-14-alkyl esters (CAS 84238-60-8), 2-Propenoic acid, C16-18-alkyl esters (CAS 90530-21-5), and 2-Propenoic acid, C18-22-alkyl esters (CAS 85085-17-2).No toxicity to bacteria was observed, the EC10 / EC50 values were, depending on the test conditions, > 100 mg/L. Thus, concentrations of the test substances were several orders of magnitude above the limit of solubility. There is, however, no indication that the undissolved test substances interfered with bacterial respiration or the functioning of the test system. In conclusion, all members of the category (C12-C22) are considered to be non-toxic to microorganisms. Hence no inhibition of biodegradability in STPs or in the environment is expected.

Key value for chemical safety assessment

EC50 for microorganisms:

1 000 mg/L

Additional information

2-Propenoic acid, C12-14-alkyl ester was tested withPseudomonas putidain accordance to DIN 8192. The test concentrations were 100, 1000 and 10000 mg/L. The EC0 was >= 10000 mg/L and the EC10 was also greater than 10000 mg/L (BASF SE 1988).

A study according to OECD No. 209 was performed to determine the effect of different concentrations of the test substance, 2-Propenoic acid, C16-18-alkyl esters (CAS 90552-04-8) on the oxygen consumption of microbial-inoculum, compared to the blank control. The test was conducted at concentrations of 62.5, 125, 250, 500 and 1000 mg/L. Based on these results, the EC50 value was calculated to be greater than 1000 mg/L (BASF SE, 2012).

 

A determination of the inhibition of oxygen consumption in the activated sludge respiration inhibition test for the test substance2-Propenoic acid, C18-22-alkyl esterswas performed. The study was conducted according to test guideline OECD Guideline 209.

The test was conducted at concentrations 1000, 500, 250, 125, 62.5 mg/L as nominal concentration based on test substance without correction of purity or composition and blank controls. Incubation time was 3 hours. Sludge concentration in the test: 1.5 g/L and the inoculum was activated sludge from a municipal wastewater treatment plant from the municipal wastewater treatment plant of Mannheim, Germany. Based on these results, the EC50 value was calculated to be greater than 1000 mg/L (BASF SE, 2011).