N-methoxy-1-(2,4,6-trichlorophenyl)propan-2-amine

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Remarks:

The acute earthworm toxicity study was conducted solely to comply with a non-EU national registration requirement, and has been provided here in accordance with REACH, Article 22(1)e.

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 April 2015 to 03 June 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

yes

Remarks:

finely ground quartz sand

Details on preparation and application of test substrate:

Since the test item has low water solubility, an exact weighed amount of the test item was mixed with finely ground quartz sand, immediately prior to application. This stock mixture was diluted with quartz sand in a way that 10 g of the mixture contained the amount of test item required for one replicate to adjust the selected concentration. The treated quartz sand (10 g per replicate) was added to the prepared amount of artificial soil (741 g wet weight) yielding 751 g wet artificial soil (corresponding to 556 g dry weight). The test item mixture was then mixed thoroughly with the artificial soil by intensive stirring in a laboratory mixer.
The order of application was as follows:
- quartz sand (untreated control)
- test item mixtures (increasing order)

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Age and weight range at test start: adult worms (approximately 3 months old with clitellum)
301 – 468 mg/worm
Source: reared at test facility (originally: “W. Neudorff GmbH KG”, An der Mühle 3, 31860 Emmerthal, Germany)
Feeding: None during test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

19.7 – 21.9 ºC

pH:

test start: 5.95 – 6.03
test end: 5.78 – 5.93

Moisture:

test start: 34.9 – 35.1 (equivalent to 56.4 – 56.7 % of WHC)
test end: 34.2 – 34.7 (equivalent to 55.3 – 56.1 % of WHC)

Details on test conditions:

Vessels: 1 L glass jars with glass lids allowing gas exchange
Substrate: Artificial soil comprising 10% sphagnum peat, 20 % kaolinite clay, 69.5 % industrial quartz sand (> 50% of the particles between 0.05 mm and 0.2 mm) and 0.5% calcium carbonate. 751 g wet weight soil, corresponding to ca. 556 g dry weight, of artificial soil was added to each test vessel
Replication: 4
No. of worms/arena : 10
Photoperiod: Continuous illumination (510 lux)

Nominal and measured concentrations:

31.25, 62.5, 125, 250, 500, 1000 mg a.i./kg soil d.w. (equivalent to 33.5, 67.1, 134, 268, 536, 1073 mg test item/kg soil d.w.)

Reference substance (positive control):

yes

Remarks:

2-chloroacetamide

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

626 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: biomass

Details on results:

At the beginning of the test, the fresh weight of the earthworms was in the range of 301 – 468 mg/earthworm. The physico-chemical parameters of the test substrate measured at the start and at the end of the test met the guideline requirements. The maximum burrowing time was 5 minutes in the control group and in the CA5204A treated groups.
Mortality rates of 0 % - 100 % were recorded in the test item treatment groups. No mortality was observed in the control group. Statistically significant mortality of 100 % was found at 1000 mg a.i./kg soil d.w. (Fisher’s Exact Binomial Test with Bonferroni Correction, p ≤ 0.05, one-sided greater). No abnormal behaviour or toxic symptoms of the worms were observed in the test item treatment groups or control group during the test.
The test item caused no statistically significant change in biomass (difference in fresh weight of surviving worms between test start and day 14 of exposure) compared to the control (7.8 % biomass decrease) at concentrations of 31.25, 62.5, 125, 250 and 500 mg a.i./kg soil d.w. (biomass decreases of 7.2, 7.1, 8.0, 7.5 and 10.9 %, respectively, were recorded). The biomass change at 1000 mg a.i./kg soil d.w. could not be determined, because all worms died at this concentration before the end of the test.

Results with reference substance (positive control):

The reference item 2-chloroacetamide was routinely tested at concentrations of 14.1, 18.3, 23.8, 31.0 and 40.3 mg a.i./kg soil d.w. In the most recent study with the reference item (BioChem project No. R 15 10 48 002 S, dated 28 April 2015), the 14-day LC50 was calculated as 21.3 mg a.i./kg soil d.w. with 95 % confidence limits ranging from 20.3 to 22.4 mg a.i./kg soil d.w. The test results indicate that the laboratory conditions are adequate and have not changed over time

Reported statistics and error estimates:

The statistical analysis was performed with the software ToxRat Professional 3.0.beta. Fisher`s Exact Binomial Test with Bonferroni Correction and the Williams-t-test were used to compare the control with the independent test item groups. The LC50 value was calculated by the Trimmed Spearman-Karber procedure. For statistical evaluation of the biomass change, the changed mean fresh weight of surviving worms per replicate was used.

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test in which earthworms (Eisenia fetida) were exposed to CA5204A the NOEC based on biomass was determined to be 500 mg a.i./kg soil d.w.
The 14-d LC50 was determined to be 626 mg a.i./kg soil d.w.

Executive summary:

CA5204A was tested in an Acute Toxicity to the Earthworm Eisenia fetida in  Artificial Soil according to OECD guideline 207.

Seven concentrations (31.25, 62.5, 125, 250, 500, 1000 mg a.i./kg soil d.w. (factor 2) (equivalent to  33.5, 67.1, 134, 268, 536, 1073 mg test item/kg soil d.w.  corrected for purity))

were tested on 4 replicated each containing 10 worms. The mortality and behaviour were assessed 7 and 14 days after application and the biomass was assessed 14 days after application.

Approximately 24 hours prior to test start, the artificial soil was prepared and deionised water was added to the dry components.The test item was introduced by dispersing the quantity of the test item to obtain the desired test concentration in a portion of deionised water necessary to wet the soil substrate to the required water content corresponding to 40-60 % of WHC. The control substrate contained the corresponding amount of corresponding amount of deionised water only. The test vessels were then filled with the treated soil. The worms were acclimatised in a separate batch of the untreated artificial substrate for approximately 24 hours before test start. The acclimatised test animals were washed, gently dried on a paper towel, weighed and randomly placed onto the test substrate.

Description of key information

In an acute toxicity test in which earthworms (Eisenia fetida) were exposed to CA5204A the NOEC based on biomass was determined to be 500 mg a.i./kg soil d.w.

The 14-d LC50 was determined to be 626 mg a.i./kg soil d.w.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

626 mg/kg soil dw