Hydrocarbons, C4-7, C5-rich, isoprene-purifn. by-product

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May to October 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: the study was performed according to OECD guideline and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

Instead of maximal 1000 mg/L the concentration range in the screening test was set at up to 100 mg/L. This represents a deviation of the study plan which is considered to have no influence on the integrity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

The values >21°C are slightly out of the temperature range set at 20 ± 1°C. This represents a deviation of the study plan which is not considered to have an influence on the integrity of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Absorbent A4

IUPAC Name:

Absorbent A4

Details on test material:

- Name of test material (as cited in study report): Absorbent A4
- Substance type: UVCB
- Physical state: Transparent liquid without mechanical impurities, colour from light to dark
- Stability under test conditions: months
- Storage condition of test material: room temperature, in the dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 12.8 mg / 2000 mL, 25.0 mg / 2000 mL, 49.9 mg / 2000 mL, 100.8 mg / 2000 mL, and 200.3 mg / 2000 mL
- Sampling method: data not available
- Sample storage conditions before analysis: data not available

Test solutions

Vehicle:

no

Details on test solutions:

The test item was tested for toxicity to¬wards daphnia as aqueous extracts (“WAF”) from up to 100 mg test item/L (final concentration). For this purpose, different amounts of the test item were dispersed in dilution water. The elution of the water soluble part (WAF) of the test item took place on a shaking machine at 130 rpm speed over 24 h at room temperature, respectively. During that time an equilibration between the test item and the water was considered to be achieved. Thereafter, the suspensions were filtered through glass fibre filters in order to eliminate non-dissolved parts of the test item. The eluates were used for the test without further dilution steps.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain: Daphnia magna STRAUS (clone 5)
- Source: Of a laboratory breed (derived from Dr. M. Bergtold, BASF, D-67117 Limburgerhof; the strain originally derived from the Institut National de Recherche Chimique Appliquee, France in 1978 and cultivated since then at BASF using medium M4 acc. Elendt) was used for the test.
- Age at study initiation: not older than 24h
- Method of breeding: the cultivation of the daphnias is performed in a way that the animals are transferred in new test medium at an interval of 1 to 3 days followed by feeding with Desmodesmus subspicatus- and/or Chlorella-cells.
- Feeding during test: during the test the daphnia were not fed

ACCLIMATION: data not available

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

none

Test conditions

Hardness:

data not available

Test temperature:

see table 1

pH:

see table 1

Dissolved oxygen:

see table 1

Salinity:

not applicable (freshwater)

Nominal and measured concentrations:

see table 2 and 3

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL carboys made of glass
- Type (delete if not applicable): data not available
- Aeration: data not available
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: data not available

TEST MEDIUM / WATER PARAMETERS: test medium according Elendt M4 according to the guideline (prepared in ultra pure water)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h of light and 8h of darkness
- Light intensity: data not available

EFFECT PARAMETERS MEASURED: after 24h and 48h the number of immobilised daphnia at each concentration of the test item is recorded

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study
- Test concentrations: 1 mg/L, 10 mg/mL and 100 mg/L
- Results used to determine the conditions for the definitive study: no immobilisation observed at the concentration of 1 mg/L

Reference substance (positive control):

no

Remarks:

A reference item was not used within the biological part of this study. Within the analytical part of the study, the test item served as the reference.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EL10

Effect conc.:

1.56 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

1.81 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.98 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

1.84 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

In order to verify the sensitivity of the test system towards toxic items the effect of potassium dichromate towards different animals of this breed was tested prior to beginning of the test. The value of the EC50 (24h) in this period of time was 1.06 mg/L potassium dichromate. This value is in accordance with the recommendations given in OECD Guideline 202 (Daphnia sp., Acute Immobilisation Test).

Reported statistics and error estimates:

not applicable

Any other information on results incl. tables

Main Test

 

Nominal Concentration of the testitem(mg/L) (Eluates/“WAF“)		Ratio immobilised Daphnia [%]		pH-Value		O2-Content [mg O2/L]		Temperature*)[°C]
		24h	48h	t0	48h	t0	48h	t0	48h
Control (0)		0 [0/20]^1)	0 [0/20]^1)	8.74	8.87	8.6	8.4	21.1	20.7
6.25		0 [0/20]^1)	0 [0/20]^1)	8.63	8.70	7.2	7.5	21.0	20.8
12.50		20 [4/20]^1)	55 [11/20]^1)	8.71	8.78	6.7	7.4	21.0	20.8
25.00		100 [20/20]^1)	100 [20/20]^1)	8.63	8.75	6.1	6.5	21.1	20.5
50.00		100 [20/20]^1)	100 [20/20]^1)	8.78	8.85	5.7	6.0	21.1	20.6
100.00		100 [20/20]^1)	100 [20/20]^1)	8.68	8.79	6.1	6.4	21.1	20.7

 

¹⁾Numbers in brackets give the absolute numbers of immobilised animals at each concentration.

 

 

Effective Concentrations on Basis of the Nominal Concentrations: 

 

	t24h	t48h
NOEC	12.5 mg/L	6.25 mg/L
LOEC	25.0 mg/L	12.5 mg/L
EC10	11.8 mg/L	10.3 mg/L
EC20	12.5 mg/L	11.0 mg/L
EC50	13.9 mg/L	12.3 mg/L

 

 

Effective Concentrations (Loadings) on Basis of the Measured Concentrations:

 

	t24h	t48h
NOEC	1.84 mg/L	0.98 mg/L
LOEC	3.74 mg/L	1.84 mg/L
EC10	1.75 mg/L	1.56 mg/L
EC20	1.84 mg/L	1.65 mg/L
EC50	2.02 mg/L	1.81 mg/L

 

*)Temperatures were partially outside the range of 20 ± 1°C indicated in the study plan. This represents a deviation of the study plan which is not considerd to have an influence on the integrity of this study.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

In the control without any test item no dead animals were observed. In the test solutions the oxygen concen¬tration did not drop below a value of 60% of the saturation value (ca. 5 mg O2/L).

Conclusions:

The reported effect concentrations refer to mean measured concentrations (geometric mean) of the measured concentrations. The following EC-values (effective concentrations) were determined acordingly:
t24h t48h
NOEC 1.84 mg/L 0.98 mg/L
LOEC 3.74 mg/L 1.84 mg/L
EC10 1.75 mg/L 1.56 mg/L
EC20 1.84 mg/L 1.65 mg/L
EC50 2.02 mg/L 1.81 mg/L

Executive summary:

”Absorbent A4“was tested for acute toxicity towards daphnia according to OECD-Test Guideline 202 (version dated 13-Apr-2004). In order to in­vestigate the influence of the test item towards the daphnias, the swimming behaviour of the animals was recorded. Under the conditions used for the test, immobilisa­tion of the daphnias was observed at concentrations representing the water soluble part (hereafter abbreviated “WAF”, water accommodated fraction) of the test item at concentrations up to 100 mg/L (screening test). As a conclusion of the analytical part of this study, recoveries of the concentrations determined after 48h were not all at ≥ 80% of those determined at time t₀, and furthermore,the measured concentrations of the treatments deviated more than 20 % from the nominal values. Based on this, the reported effect concentrations refer tomean measuredconcentrations (geometric mean) of the measured concentrations.The following EC-values (effective concentrations) were determined acordingly:

 

	t24h	t48h
NOEC	1.84 mg/L	0.98 mg/L
LOEC	3.74 mg/L	1.84 mg/L
EC10	1.75 mg/L	1.56 mg/L
EC20	1.84 mg/L	1.65 mg/L
EC50	2.02 mg/L	1.81 mg/L

 

The test is considered valid as all conditions for validity were met.

 

·        In the control without any test item no dead animals were observed.

·         In the test solutions the oxygen concen­tration did not drop below a value of 60% of the saturation value (»5 mg O₂/L).