1-(N,N-bis(2-ethylhexyl)aminomethyl)-1,2,4-triazole

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Remarks:

Degradation product

Adequacy of study:

key study

Study period:

exposure period: 23 Aug 2012 to 17 Oct 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
As the substance hydrolyses immediately in contact with water, data from the major constituent of the hydrolysate (bis(2 -ethylhexyl)amine is used to assess this endpoint. This constituent is also the most hazardous constituent to the aquatic environment. For further read across justification see IUCLID chapter 6.1 and CSR chapter 7.1.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Parent substance EC 401 -280 -0
The acute toxicity of the substance EC 401-280-0 to Daphnia was assessed according to OECD guideline no. 202 (1984). The respective EC50 24 h is 9.3 mg/L, based on nominal concentrations. As the exposure was only 24 hours, this data is only used as supportive information.

Furthermore, as the substance hydrolyses immediately in contact with water, data from the major constituent of the hydrolysate (bis(2 -ethylhexyl)amine is used to assess this endpoint. This constituent is also the most hazardous constituent to the aquatic environment. For further read across justification see IUCLID chapter 6.1 and CSR chapter 7.1.

Hydrolysis products
bis(2 -ethylhexyl)amine
The reported 48-h EC50 of bis(2-ethylhexyl)amine on Daphnia magna was 2.2 mg/L (nominal value) in a test using Tween 80 as a solvent. The EC50 was determined using non-neutralised samples (BASF AG 1/0500/2/89-0500/89, 1989).
Based on the results from a long-term reproduction study with Daphnia magna, the substance is classified as Chronic Category 1. Comparing the available aquatic toxicity data on the substance EC 401 -280 -0 with the data of bis(2 -ethylhexyl)amine shows that aquatic toxicity is in the same range. In most cases the derived effect concentrations of bis(ethyhexyl)amine are lower.Therefore, aquatic toxicity data of the major and the most hazardous component of the hydrolysate (bis(2-ethylhexyl)amine) is used as worst case to assess the acute toxicity to aquatic invertebrates of the substance EC 401-280-0 and its hydrolysate.

3. ANALOGUE APPROACH JUSTIFICATION
The aquatic toxicity data of the major and the most hazardous component of the hydrolysate (bis(2-ethylhexyl)amine) is used as worst case to assess the acute toxicity to aquatic invertebrates of the substance EC 401-280-0 and its hydrolysate.

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): The test item was dissolved in a sufficient volume of acetone to prepare a stock solution. This stock solution was dissolved with acetone to obtain one test item application solution (= dosage solution) for each test item concentration in such a way that an appropriate amount of these test item/acetone solutions were used to soak the 10% portion of the quartz sand. The solvent was evaporated to retrievethe coated quartz sand fractions, which were incorporated into the remainders of the artificial soil to prepare the different test item soil concentrations.
- Controls: solvent treated, water treated
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution): test item dissolve in pure acetone (no water added)
- Evaporation of vehicle before use: yes

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: earthworms kept at test facility since Feb. 1994 (original source: Co. Landenberger (Schömberg, Germany)
- Age at test initiation (mean and range, SD): adult worms with clitellum, age of individuals did not differ by more than four weeks, range: >= 2 months, < 1 year
- Weight at test initiation (mean and range, SD): mg fresh weight: control: 392.1±27.1, solvent control 390.5±32.4

ACCLIMATION
- Acclimation period: one day
- Acclimation conditions (same as test or not): yes (acclimatised in artificial soil)
- Health during acclimation (any mortality observed):

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Remarks:

adult worms removed from substrate after 28 d

Test temperature:

18.3 - 21.9 °C

pH:

day 0: 6.1 – 6.2
day 56: 6.7 – 6.8

Moisture:

day 0: 47.5 – 52.1% WHCmax
day 56: 53.0 – 58.1% WHCmax

Details on test conditions:

TEST SYSTEM
- Test container (material, size):
- Amount of soil or substrate:
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): artificial soil according to OECD 222
- 5% Sphagnum peat (no visible plant remains; finely ground and air-dried)
- 20% Kaolin-Clay (kaolinite content > 30%)
- 74-75% Quartz sand (fine sand content with particles between 50 and 200 µm higher than 50%)
- 0.3-1% Calcium carbonate (CaCO3)
- Maximum water holding capacity (in % dry weigth): 65.7%

OTHER TEST CONDITIONS
- Photoperiod: light/dark : 16 h/8 h
- Light intensity: 417 - 459 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study: performed (no data provided in study report)

Nominal and measured concentrations:

nominal: 5, 10, 20, 40 and 80 mg test item/kg artificial soil (dw); solvent control, water control

Reference substance (positive control):

yes

Remarks:

carbendazim (as active ingredient in Derosal)

Key result

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

20 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

56 d

Dose descriptor:

LOEC

Effect conc.:

40 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

56 d

Dose descriptor:

EC50

Effect conc.:

> 80 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 80 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Remarks:

(biomass)

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

> 80 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Remarks:

(biomass)

Details on results:

- Mortality at end of exposure period: solvent control: 0%
- Total mass of adults at beginning of test:
- Changes in body weigth of live adults (% of initial weight) at end of exposure period:
- No. of offspring produced: solvent control: 525±52.9
- Morphological abnormalities: no
- Behavioural abnormalities: no
- Other biological observations: Almost the whole food was consumed by the worms in all concentrations after 7, 14, 21 and 28 days of the application of the test item.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: NOEC reproduction = 0.32 mg/kg soil (dw); LOEC reproduction = 1.0 mg/kg soil (dw)

Reported statistics and error estimates:

The statistical software package ToxRat Professional 2.10 was used for the statistical evaluation of reproduction and biomass. Statistical evaluation of mortality was not performed due to 0% mortality of the adult earthworms.

Physical parameters

Concentration [mg test item/kg soil (dw)]	pH		pH
	Start	End	Start		End
			% of dry mass	% of WHCmax	% of dry mass	% of WHCmax
Control	6.2	6.8	32.5	49.5	34.8	53.0
Solvent control	6.2	6.7	31.6	48.1	36.8	56.0
5	6.2	6.7	32.5	49.5	36.8	56.0
10	6.1	6.7	32.5	49.5	35.9	54.6
20	6.2	6.7	32.1	48.9	36.3	55.3
40	6.2	6.8	31.2	47.5	38.2	58.1
80	6.1	6.7	34.2	52.1	34.8	53.0

Biomass of the earthworms at test start and after 28 d of exposure

Concentration [mg test item/kg soil (dw)]	Biomass (mg fresh weight)		Percent of the initial weight
	Day 0	Day 28	Day 28
Control	392.1±27.1	503.6±25.5	128.4
Solvent Control	390.5 ± 32.4	547.0 ± 19.3	140.1
5	393.3 ± 25.9	550.7 ± 31.8	140.0
10	395.3 ± 20.7	554.1 ± 28.3	140.2
20	391.8 ± 20.8	548.6 ± 20.2	140.0
40	388.6 ± 25.5	563.2 ± 27.0	144.9
80	393.0 ± 18.3	551.2 ± 20.7	140.3

Number of juveniles after 56 d

Concentration [mg test item/kg soil (dw)]	Number of Juveniles per Treatment (mean ± sd)	Number of Juveniles [% of Solvent Control]
Control	567.3 ± 30.5	108.1
Solvent Control	525.0 ± 52.9	100.0
5	469.0 ± 51.7	89.3
10	561.0 ± 75.5	106.9
20	526.5 ± 58.2	100.3
40	446.3 ± 52.2	85.0
80	317.0 ± 31.1	60.4

Effects on Mortality, Biomass and Reproduction

Concentration [mg test item/kg soil (dw)]	Mortality [%]	Biomass [% of initial weight]	Juveniles [% of Control]
	Day 28	Day 28	Day 56
Control	0.0	128.4	108.1
Solvent Control	0.0	140.1	100.0
5	0.0	140.0	89.3
10	0.0	140.2	106.9
20	0.0	140.0	100.3
40	0.0	144.9	85.0*
80	0.0	0.0	60.4*

*: Significantly different to control (Williams t-test; 1 -sided, p <= 0.05).

Validity criteria fulfilled:

yes

Conclusions:

The 56d-NOEC reproduction of Eisenia fetida exposed to bis(2-ethylhexyl)amine was determined to be 20 mg/kg (dw).

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Description of key information

The 56d-NOEC reproduction of Eisenia fetida exposed to bis(2-ethylhexyl)amine was determined to be 20 mg/kg (dw). This value is used for the assessment of terrestrial toxicity of substance EC 402-280-0 and its hydrolysate.

Key value for chemical safety assessment

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

20 mg/kg soil dw

Additional information

Parent substance EC 401-280-0

No data on terrestrial toxicity are available.

As the substance hydrolyses immediately in contact with water, data from the major constituent of the hydrolysate (bis(2 -ethylhexyl)amine is used to assess this endpoint. This constituent is also the most hazardous constituent to the aquatic environment. For further read across justification see IUCLID chapter 6.1 and CSR chapter 7.1.

Up to Annex VIII of the REACH Regulation (EC) No 1907/2006 this information is not required. According to Annex IX, studies on the effects on terrestrial organisms shall be conducted for substances manufactured or imported in quantities of 100 tons or more.

The risk characterization ratios (RCRs) of the chemical safety assessment for substance EC 401-280-0 are below 1, indicating that the risks are acceptable. Therefore, no studies on terrestrial organisms other than earthworms are currently provided. Since the major an most hazardous constituent of the hydrolysate (2-bis(ethylhexyl)amine) has a high potential to adsorb to soil particles and is not readily biodegradable, available data on long term reproduction study on earthworms with bis(2-ethylhexyl)amine are regarded as sufficient to assess terrestrial toxicity of substance EC 401 -280 -0 and its hydrolysate.

 

Hydrolysis products:

Bis(2-ethylhexyl)amine

A long term reproduction study on earthworms according to OECD 222 is available (BASF SE and OXEA Group, 2012). Test substance concentrations up to 80mg/kg (dw) were tested.

The substrate was artificial soil. After 28 days of exposure no mortality was observed for the adult earthworms. The effect on reproduction and biomass was assessed after a further 28 days. The effect values for biomass were as follows: NOEC >= 80 mg/kg soil dw; LOEC > 80 mg/kg soil dw. The effect values for reproduction were: NOEC = 20 mg/kg soil dw; LOEC = 40 mg/kg soil dw.

 

Formaldehyde

No data on guideline studies on soil toxicity are available. As formaldehyde is readily biodegradable, not classified for environmental hazards according to CLP regulation and is only a minor constituent of the hydrolysate, data on soil toxicity are not considered to be relevant for the assessment of terrestrial toxicity of substance EC 401-280-0 and its hydrolysate.

 

1H-1,2,4-triazole

Supporting data on acute toxicity of 1H-1,2,4-triazole to to earthworms are available. The test was performed according to OECD guideline 207(ECHA, 2014). No mortalities were observed even at the highest test concentration (1000 mg/kg dry weight substrate). LC50 14(d) was determined to be > 1000 mg/kg (bw).