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REACH

Thiophanate-methyl

EC number: 245-740-7 | CAS number: 23564-05-8

Life Cycle description
Uses advised against

Ecotoxicological information

Toxicity to soil microorganisms

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil microorganisms

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-08-26 to 2012-01-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 216 (Soil Microorganisms: Nitrogen Transformation Test)

Version / remarks:

2000

Deviations:

Qualifier:

according to guideline

Guideline:

OECD Guideline 217 (Soil Microorganisms: Carbon Transformation Test)

Version / remarks:

2000

Deviations:

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

Vehicle:

Details on preparation and application of test substrate:

AMENDMENT OF SOIL
One common agricultural soil was used for the study. The sampling depth was 0 - 20 cm (mixed sample). The soil parameters were in accordance with the recommendations of the OECD guidelines 216 and 217.
The test soil was sieved to a particle size of 2 mm. Prior to the initiation of the study, the moisture content of the soil and the amount of water needed to adjust the soil moisture content to 45 % WHCmax (maximum water holding capacity) were calculated. The test item was applied at a concentration of 7.0 and 35.0 mg/kg soil dry weight, respectively.
For the nitrogen turnover the soil and finely ground lucerne meal (source: Obere Muhle, D-75399 Kapfenhardt; sieve size < 1 mm, C/N-ratio: 15:1) were thoroughly mixed before application by hand. The final concentration of the dried lucerne meal was 0.5 % of the soil dry weight.
For the short-term respiration the amount of glucose needed to obtain maximal short-term rates of respiration in the test soil was determined prior to the beginning of the test. A concentration of 200 mg glucose/100 g soil wet weight was found to be the optimum and was used in the test.

APPLICATION OF TEST SUBSTANCE TO SOIL
- Method: For application, 5 mL of the test solutions were transferred onto 29.1 g quartz sand. After evaporation of the solvent the quartz sand was mixed with the appropriate amount of moist soil (3300 g) to obtain the assigned nominal test concentrations for the test vessels. Accordingly solvent treated quartz sand and untreated quartz sand were mixed with the appropriate amount of soil for the control treatments. To achieve 45 % of WHCmax 87.4 g deionised water was added to each treatment group. Each treatment group (soil with the test item added at two different concentrations, solvent control or untreated control) contained 3300 g moist soil for the nitrogen turnover and for the short-term respiration.

Test organisms (inoculum):

soil

Total exposure duration:

28 d

Test temperature:

20.5 - 21.5 °C

Moisture:

36.40 WHCmax (%)

Organic carbon content (% dry weight):

1.34

Nitrogen content (% dry weight):

0.12

Details on test conditions:

TEST SYSTEM
- Test container: glass bottles
- Amount of soil: approx. 1100 g

SOURCE AND PROPERTIES OF SUBSTRATE
- Geographical reference of sampling site (latitude, longitude): UTM Zone 32 E: 440939, N: 5449795
- History of site: The sampling site was not cultivated since 2007. For at least five years prior to soil sampling no plant protection products had been applied to the sampling site. No organic fertilizer had been applied to the site for at least six months prior to soil sampling. No mineral fertilizer had been applied to the site for four years prior to soil sampling.
- Depth of sampling:
- Soil texture (DIN): medium loamy sand
- % sand: 63.9
- % silt: 27.5
- % clay: 8.6
- pH (in solvent control): 6.77
- Initial nitrate concentration for nitrogen transformation test (mg nitrate/kg dry weight): 17.8
- Maximum water holding capacity (in % dry weigth): 36.40
- Cation exchange capacity (meq/100 g): 10.7
- Storage (condition, duration): stored at approx. 4 ± 2 °C
- Initial microbial biomass as % of total organic C: 11.6

Nominal and measured concentrations:

nominal concentrations: 7.0 and 35.0 mg/kg

Reference substance (positive control):

yes

Remarks:

Sodium Chloride

Key result

Duration:

28 d

Dose descriptor:

ER50

Effect conc.:

> 35 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

respiration rate

Remarks on result:

other:

Remarks:

no adverse effect on soil respiration

Key result

Duration:

28 d

Dose descriptor:

ER50

Effect conc.:

> 35 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

nitrate formation rate

Remarks on result:

other:

Remarks:

No adverse effect on nitrogen turnover

Results with reference substance (positive control):

The toxic reference item (Sodium Chloride), tested in a separate study had significant effects on the soil nitrogen turnover (decrease of the nitrogen level of 43.5 %, decrease of the nitrate transformation rate for the interval of 0 to 28 days of 106 %, decrease of the nitrate transformation rate for the interval of 14 to 28 days of 102 %) and the short-term respiration (94.1 % inhibition) in a field soil tested at a concentration of 20.0 g/kg soil dry weight.

Reported statistics and error estimates:

The results of the nitrification (nitrate content at day 28, nitrate formation rate of treated soils between the 14 day sampling and the 28 day sampling and nitrate formation rate over the whole study period (0 day sampling to 28 day sampling) and short-term respiration measurements (last sampling: 28 days) were tested for normality using Shapiro-Wilk's test and residual analysis (ZAR, 1999). Homogeneity of variances was tested using Levene's test or Bartlett's test. The control treatments were tested for significant differences using t-Test, pooled. All statistical evaluations were performed compared to the solvent control. For the nitrogen turnover (nitrate content at day 28, nitrate-N formation rate of the last sampling interval (14 day sampling to 28 day sampling), for the nitrate-N formation rate of the whole study period (0 day sampling to 28 day sampling) and for the short-term respiration the data comply with the requirements for a multiple-t-test. Hence the Dunnett's t-Test was used to analyse the data for significance. The statistical software program SAS version 9.2 (Ed. 2002 - 2008) was used for the statistical analysis. The individual calculation procedures are part of the raw data. For the statistical analysis of the nitrogen turnover the nitrate formation rate values of each replicate were used, but reported nitrate values per day are calculated from the means of the amount of nitrate-N.

Table 1: Test item solutions

Solution	Test concentration [mg test item/kg]	Test concentration [mg/kg sdw]	Test item/ solution (required)	Test item/ solution (weighed)	Acetone added to total amount [mL]	Deionised water added [g]	Application amount [g]
TG2	35.0	35.526	414 mg test item	414 mg test item	20.0	87.4	5¹⁾
TG1	7.0	7.105	4.0 mL TG2	4.0 mL TG2	20.0	87.4	5¹⁾
Solvent Control	-	-	-	-	-	87.4	5^{2 )}
Control	-	-	-	-	-	87.4	-

sdw = soil dry weight

¹⁾ the solutions were applied with 29.1 g quartz sand to 3300 g soil (2910.6 g soil dry

weight)

²⁾ pure acetone

Table 2: Amount of nitrate-N in control and soil treated with the test item

Days	Solvent Control	Control		7.0 mg a.i./kg soil dry weight		35.0 mg a.i/kg soil dry weight
after applica tion	NO₃^--N [mg/kg soil dry weight]	NO₃^--N [mg/kg soil dry weight]	Deviation from solvent control [%]¹	NO₃^- -N [mg/kg soil dry weight]	Deviation from solvent control [%]¹	NO₃^--N [mg/kg soil dry weight]	Deviation from solvent control [%]¹
0	17.8	18.4	3.37	18.5	3.93	18.4	3.37
7	13.7	14.5	5.84	14.4	5.11	14.6	6.57
14	24.4	25.1	2.87	27.3	11.9	28.7	17.6
28	44.9	44.6	-0.668	48.5	8.02	51.9*	15.6