2-hydroxy-1-(4-(4-(2-hydroxy-2-methylpropionyl)benzyl)phenyl)-2-methylpropan-1-one

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.8 (Toxicity for Earthworms: Artificial Soil Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

Based on the specified water solubility the application of the test substance via an aqueous stock solution is not possible. The water solubility is not enough to prepare a sufficient stock solution with deionized water.
Application of the test substance: distributed in quartz sand and added to the test substrate afterwards.
The required amounts of test substance were weighed into a beaker and mixed with 25 g quartz sand.
For preparation of the test assays with test substance amounts of about 2538 g dry test substrate (depend on the water content of the dry test substrate) will be added to a box. The quartz sand aliquots treated with the required test substance amounts will be added and thoroughly mixed with the test substrate. Than the required volumes of 713 mL deionized water will be added to the test substrate test substance mixtures and mixed well again. Four portions of approx. 750 g of the treated wet test substrate for each test concentration will be distributed to preserving jars.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earthworm Eisenia Fetida
- Source: Agricultural Center Limburgerhof of BASF SE, 67056 Ludwigshafen, Germany
- Age at test initiation (mean and range, SD): approx. 10 months (guaranteed by the supplier)
- Weight at test initiation (mean and range, SD): 303 – 599 mg (and a clear visible clitellum)
- Conditions of storage at the test facility: The worms were stored in the transport box until adaptation at room temperature in the dark.
- Adaptation: Stored in test substrate at test conditions one day before start of exposure
- Reason for selection of species: Recommended species in the Test guidelines, its susceptibility to chemicals resembles that of true soil inhabiting species, it has a short life cycle, hatches from cocoons in 3 to 4 weeks and reaches maturity in seven to eight weeks at 20 °C

ACCLIMATION
A transport box containing approx. 1950 g moistened test substrate with a content of water of about 30 % was provided to the supplier.
One day before start of exposure a sufficient number of worms were sorted out by the supplier from the breeding box and placed in the transport box with the prepared test substrate. For acclimatization to the prepared test substrate and to the test temperature the transport box was stored in the incubation room for one day under the temperature of 20 ± 2°C and continuous light without feeding.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

21.2 – 21.4 °C

pH:

5.9 - 6.1

Moisture:

Calculated water content of the test substrate at start of exposure: 30 % of dry weight

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Preserving jars, volume 1 L, covered with glass lids
- Amount of soil or substrate: 750 g ± 1 g artificial soil (wet weight)
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per refernce substance: 4

TEST SUBSTRATE
Actual mixture:
- 22.4 kg quartz sand (69.8 % of total test substrate mass)
Supplier: Gebrüder Willersinn GmbH, 67071 Ludwigshafen, Germany
- 6.4 kg kaolin clay (19.9 % of total test substrate mass)
Supplier: Co. Bassermann minerals GmbH, 68169 Mannheim, Germany
- 3.2 kg sphagnum peat (10.0 % of total test substrate mass), finely grounded
Supplier: Raiffeisen Mannheim eG, 68307 Mannheim, Germany
- 90.1 g CaCO3, p.A. (0.2 % of total test substrate mass)
Supplier: Merck
Total mass of the test substrate: 32.1 kg

To determine the water content of the dry artificial soil two parallels with about 11 g of the test substrate were weighed on a moisture determination balance. The substrate was dried at 105 ± 5 °C to constant weight. The water content was calculated from the difference in weight of substrate before and after the drying procedure.
Water content of the prepared test substrate: 1.5 % (mean value)
To determine the pH value 2 replicates with about 10 g dry weight of test substrate were weighed in a beaker. Then 50 mL deionized water was added. This mixture was stirred for 5 minutes and rested for 2 hours. After shaking the pH value was measured in the settling sample.
pH-values in the prepared test substrate: 6.2 (mean value)

OTHER TEST CONDITIONS
- Photoperiod: Continuously, over the exposure period
- Light intensity (at start of the exposure): 619 Lux, mean value of 5 single values (760, 541, 531, 660, 603 Lux)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
On day 7 and 14 the worms were removed carefully from their test vessels. During this process, morphological and behavioral changes and the number of surviving worms in each test container were recorded.
Worms were classified as dead when they did not respond to a gentle mechanical stimulus to the front end. Afterwards the worms were put back onto the soil surface in the containers. On day 14 additionally the single weight of the survived worms was determined and compared with the weights of the worms from start of exposure. The pH values of the test substrate of each test concentration were determined at start of exposure in a sample taken from the mixing bowl and in one replicate of each test concentration at the end of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Test concentrations: 0 (control), 62.5, 125, 250, 500, 1000 mg/kg soil dry weight

Nominal and measured concentrations:

62.5, 125, 250, 500, 1000 mg/kg soil dry weight as nominal concentrations based on test substance

Reference substance (positive control):

yes

Remarks:

2-Chloroacetamide

Duration:

14 d

Dose descriptor:

LC10

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

No abnormal behaviour or toxic symptoms of the worms were observed in the test substance groups or control group during the exposure

Results with reference substance (positive control):

The LC50 value of 2-Chloroacetamide for the earthworm Eisenia fetida after 14 days was 21.5 mg/kg and therefore within the required range of 20 mg/kg to 80 mg/kg. The value shows that the test system was sufficiently sensitive and suitable to determine the acute toxicity of the test substance based on the test guideline.

Effects of test substance on the earthworm (Eisenia fetida) biomass change at the end of exposure

Test substance [mg/kg soil dry weight] nominal value	0 (Control)	62.5	125	250	500	1000
Biomass change [%]	-4.5	-0.9	-6.4	-4.1	-4.4	-6.6

Validity criteria fulfilled:

yes

Conclusions:

No toxic effects have been observed in this study with soil living organisms.

Executive summary:

In a GLP-certified acute toxicity test, performed in accordance with OECD guideline 207, Eisenia Fetida were exposed to the test substance (EC 444-860-9) for 14 days. In this study, batches of ten earthworms were exposed to nominal concentrations of 62.5, 125, 250, 500, 1000 mg test substance per kg of artificial soil dry weight. A positive control of 2-chloroacetamide was used to assess the suitability of the test system. The morphological and behavioural changes were assessed at days 7 and 14. On day 14 the weight of the worms was determined and compared to the weight before exposure. In addition the pH values of the test substrate at each test concentration was determine at the start of exposure and from each test concentration at the end of exposure. The 2-chloroacetamide gave a 14-day LC50 = 21.5 mg/kg artificial soil dry weight (between the required range of 20-80 mg/kg) thus proving the test system was sufficiently sensitive and suitable to determine the toxicity of the test substance. No abnormal behaviour or toxic symptoms of the worms was observed in either test substance group or the control group, therefore the LC0 > 1000.0 mg/kg soil dw and the LC50 > 1000.0 mg/kg soil dw.

Description of key information

A GLP guideline study according OECD 207 has been conducted (BASF SE 2014). The acute toxicity of the test substance to the earthworm Eisenia fetida has been tested. After 14 d of exposure a LC50 of > 1000 mg/kg soil dw was determined. No abnormal behaviour or toxic symptoms of the surviving worms were observed in the test substance groups or control group during the exposure.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw

Long-term EC10, LC10 or NOEC for soil macroorganisms:

1 000 mg/kg soil dw

Additional information

Study conducted to recognised testing guidelines with GLP certification.