Bis(2-propylheptyl) phthalate

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-10-09 - 2018-02-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

Version / remarks:

29 July 2016

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Bis(2-propylheptyl) phthalate

EC Number:

258-469-4

EC Name:

Bis(2-propylheptyl) phthalate

Cas Number:

53306-54-0

Molecular formula:

C28H46O4

IUPAC Name:

1,2-bis(2-propylheptyl) benzene-1,2-dicarboxylate

Details on test material:

- Name of test material (as cited in study report): Palatinol 10-P
- Analytical purity: 98.3 area-% (GC, DB-1), 98.8 area-% (GC, DB-5 HT), see report No 17L00129
- Homogeneity: given
- Lot/batch No.: F151_2017
- Expiration date of the lot/batch: 07 Dec 2018

Sampling and analysis

Analytical monitoring:

no

Test substrate

Vehicle:

yes

Remarks:

aceton

Details on preparation and application of test substrate:

Palatinol 10-P is liquid but poorly soluble in water. Therefore, a stock solution with a sufficient organic solvent was prepared. The test substance was dissolved in acetone to
make a stock solution. Aliquots of this stock solution with required amounts of test substance for each scheduled test concentration were carefully mixed with quartz sand.
The organic solvent was removed by evaporation and subsequently blended with prepared test substrate.
The water solubility was not enough to prepare a sufficient stock solution of the test substance with deionized water.
Application of the test substance: dissolved in acetone
An amount of 12501.3 mg test substance was weighed into a 50 mL volumetric flask.
The test substance was dissolved by addition of acetone as solvent. The solution was filled up to the mark afterwards. The test substance was completely dissolved. The solution was colourless and clear.
The required aliquots of the stock solution were added to 25 g quartz sand (10 g quartz sand/kg dry weight). The mixture was blended thoroughly, and the organic solvent was removed by evaporation afterwards.
No odour of the organic solvent was noticeable.
The required amounts of artificial soil were weighed into a mixing tray. The test substance / quartz sand mixtures were added and thoroughly mixed with the soil. Thereafter, the water content of the soil was adjusted to about 50% WHCmax using deionized water and was mixed once again. The test mixtures were distributed to the test units.

Each replicate container was filled with about 664 g of the prepared test mixture (corresponding to about 550 g Dw artificial soil) to a layer of about 4 cm and a corresponding surface area of about 170 cm2.

Test organisms

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Breeding facility: BASF SE, Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany
Only adult worms (with clitellum) with a fresh weight between 300 and 600 mg were used.The worms that were introduced into the test were bred in horse manure and had an age from about 12 months.
Acclimatization: Before starting the test, the worms selected were acclimatized in the test substrate in a plastic box under test conditions through overnight under permanent light and without
feeding. For this, a plastic box was filled up with test substrate (750 g dry weight) and added with a sufficient number of adult worms that were sorted out from the breeding boxes.

Study design

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Test conditions

Test temperature:

20.7 °C – 21.5 °C

pH:

value of the dry test substrate: 5.9 (mean value of 2 single values; 5.8/5.9)

Moisture:

Water content in the test: 46.8% WHCmax (mean value)

Details on test conditions:

TEST SYSTEM
- Test container: Plastic dishes with a volume of about 1.0 L, dimensions 11×15.5×6 [cm] with transparent and punctured lid
- Amount of soil or substrate: Approx. 664 g artificial soil (wet weight)
- No. of organisms per container: 10
- No. of replicates per treatment group: 4 test vessels per concentration
- No. of replicates per control: 4
- No. of replicates per solvent control: 8

SOURCE AND PROPERTIES OF SUBSTRATE
- Composition (if artificial substrate): According to guideline
- Determination of water content of the dry test substrate: 0.7 % (mean of 2 single values: 0.7%/0.7%)

OTHER TEST CONDITIONS
- Photoperiod: Light/dark cycle of 16:8 hours, over the exposure period
- Light intensity: 554 Lux, (mean of 5 single measurements: 748, 496, 534, 503 and 487 [Lux])
EFFECT PARAMETERS MEASURED: Mortality, change in biomass, number of juveniles

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

The adult worms were fed with finely ground cow manure (free of growth promoters, nematicides or similar veterinary pharmaceuticals, provided by Jesus Bruderschaft e.V.
Gnadenthal, D-65597 Hünfelden). The experiment began with the application of the test substance as well as the introduction of the adult worms. Initial food supply was given
one day after this. Thereafter, the adult worms were fed weekly during the initial 4 weeks of the test. An amount of 2-5 g finely ground cow manure moistened with about 5-6 mL
deionized water was sprayed on the test substrate surface of each test container. If food remained unconsumed, the ration was reduced on demand. After removing the adults on
day 28, further 5 g of food moistened with about 5-6 mL deionized water was mixed with the test substrate of each test vessel.

Nominal and measured concentrations:

Based on a range-finder test, the following test substance concentrations were selected:
Nominal concentrations: 0 (control), 0 (solvent control), 62.5, 125, 250, 500 and 1000 mg/kg dw

Reference substance (positive control):

yes

Remarks:

Carbendazim

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Body weight change and reproduction

Effect concentrations 2

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 3

Duration:

56 d

Dose descriptor:

LOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 4

Duration:

56 d

Dose descriptor:

LOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Body weight change and reproduction

Details on results:

The mortality and the body weight change of the adult worms were observed and recorded on day 28. Reproduction behaviour measured as the number of juveniles per
test vessel after test substance application was observed on the 56th day of exposure.

No mortality of adult worms was observed in the controls and at all replicates with test substance.
In this study, we have observed a significant toxic effect only at the highest tested concentration group i.e., 1000 mg/kg dry weight of soil which is not enough to plot a dose-response curve. Therefore, to derive a
reliable dose-response relationship, a study design with more concentrations in the ranges where toxic effects are observed is recommended.

Adult worms:
Mortality: No mortality of adult worms was observed in the controls and at all replicates with test substance
NOECMortality: ≥ 1000 mg test substance/kg soil (dw)
LOECMortality: > 1000 mg test substance /kg soil (dw)

Body weight change:
NOECBody weight change: = 500 mg test substance/kg soil (dw)
LOECBodyweight change: = 1000 mg test substance /kg soil (dw)

Juvenile worms:
Reproduction by count of juvenile worms:
NOECReproduction: = 500 mg test substance/kg soil (dw)
LOECReproduction: = 1000 mg test substance/kg soil (dw)

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: EC50 = 1.02 mg/kg dw (LCL: 0.99 UCL: 1.05)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes