Reaction mass of decamethyltetrasiloxane; dodecamethylpentasiloxane; hexamethyldisiloxane; octamethyltrisiloxane

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

Adequacy of study:

weight of evidence

Study period:

22 April to 18 June 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Well conducted study according to OECD 222 and difficult substances guidance, with GLP and analysis of media.

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Natural sandy loam used, as opposed to artificial soil

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Nominal: 0.0076, 0.076, 0.76, 7.6 and 76 mg/kg
- Sampling method: During the definitive exposure, soil samples were removed on days 0, 2, 28 (before and after adults were removed) and 56. An additional test vessel was sacrificed at each interval and the soil sample analyzed for D5 concentration as to leave the test organisms and offspring undamaged. Cores were used for the determination of test concentration and the concentration of each sample was based on the mean of the three individual results
- Sample storage conditions before analysis: Directly after sampling, the cores were placed in glass bottles with teflon-lined caps and were presented for analytical processing.

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): D5 was applied directly to natural sandy loam (see Protocol Deviations) with a percent moisture of 2.9%. The soil aliquots were contained in glass jars . The soil was allowed to mix thoroughly at a speed of 15 rpm overnight on a rolling table. Table 1 describes the application of D5 to the natural sandy loam. Due to the limitations of the amount of D5 that could be directly dispensed to the soil, and the volume of the mixing jars, the lower concentrations (0.076 and 0.0076 mg/kg) were prepared by one dilution of treated soil. Table 2 describes the soil dilutions from a 7.6 mg/kg soil concentration prepared the previous day. The treated sandy loam was allowed to mix thoroughly at a speed of 15 rpm overnight on a rolling table. After treated sandy loam was mixed overnight and added to replicate vessels, 200 mL of additional deionized water was added to each replicate to bring the percent moisture to approximately 24%.
- Controls: Sandy loam without test material

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earthworm
- Eisenia fetida
- Source: Original Source of Culture: Happy D Ranch, Visalia, California
- Age at test initiation (mean and range, SD): F0 Generation age: 12 months old
- Weight at test initiation (mean and range, SD): mean individual weight ranged from 0.3855 to 0.4492 g

ACCLIMATION
- Acclimation period: One day
- Acclimation conditions (same as test or not): yes
- Health during acclimation (any mortality observed): No mortality reported

Study type:

laboratory study

Substrate type:

other: natural sandy loam

Limit test:

no

Total exposure duration:

56 d

Remarks:

28-d F0 generation phase terminated

Test temperature:

19 to 23 °C

pH:

Day 0 pH: 6.92-7.55
Day 56 pH: 6.24-6.70

Moisture:

Day 0: 11-22% (Due to the volatility of the test material, soils were not mixed after adding water to the required % moisture and do not represent the actual % moisture of the entire vessel until the water disperses evenly.)
Day 56: 19-28%

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Glass, 1000 ml
- Amount of soil or substrate: 1000 g sandy loam
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: One replicate for the 0.0076, 0.076, 0.76 and 7.6 mg/kg concentrations, eight replicates for the76 mg/kg concentration.
- No. of replicates per control: 8
Each test vessel was covered with perforated transparent plastic wrap and secured with a rubber band.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil) : Periodic analysis of representative samples of the deionized water source, natural sandy loam and cattle manure used during the earthworm culture and testing were analyzed for the presence of pesticides, PCBs and toxic metals by GeoLabs, Inc., Braintree, Massachusetts. Results of these analyses established that the deionized water source, manure and natural sandy loam did not contain any contaminants which would adversely affect the test organisms.
- Geographic location: Not reported
- Pesticide use history at the collection site: Not reported
- Collection procedures: Not reported
- Sampling depth (cm): Not reported
- Soil texture (if natural soil) Sandy loam
- % sand: 75
- % silt: 19
- % clay: 6
- Soil taxonomic classification: Not reported
- Soil classification system: Not reported
- Composition (if artificial substrate): n/a
- Organic carbon (%): The organic matter content of the soil was 5.2%. Awaiting data on OC
- Maximum water holding capacity (in % dry weigth): maximum sorption concentration: 76 mg D5/kg dry soil
- CEC:
- Pretreatment of soil: not reported
- Storage (condition, duration): not reported
- Stability and homogeneity of test material in the medium: Results of the replicate samples were generally consistent indicating homogeneous distribution of the test substance in the exposure vessels.
Losses of the test substance over time were not expected in this exposure system based on previous data collected from samples of treated natural sandy loam. Earthworm metabolism, other biological components associated with the earthworms or microorganisms associated with the manure included as an earthworm food source could have contributed to the concentration decline. Due to these factors, the biological results are reported in terms of nominal concentrations and as geometric mean measured concentrations.


OTHER TEST CONDITIONS
- Photoperiod: 16 hours of light and 8-hour dark photoperiod
- Light intensity: 340 to 750 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations on food consumption were recorded weekly, beginning on day 7, until test day 28. The group weight was determined for earthworms (F0) added to each replicate vessel at test initiation (day 0), and the group weight was determined at the termination of the F0 generation phase (day 28). Earthworm mortality and health (e.g., color changes, lethargy and softness) assessments were also performed on day 28. Any earthworms that were unaccounted for were presumed dead and decomposed.
On day 28, all cocoons were left in the test soil to continue the F1 generation exposure. At test termination (day 56 and again on day 57), the number of juveniles was determined.
pH of each soil batch was determined at test initiation and on day 56 in each replicate test vessel.
Light intensity was measured at the surface of the test vessels on days 0, 7, 14, 28, 42 and 56.
Air temperature inside the environmental chamber which housed the test vessels was monitored continuously.

VEHICLE CONTROL PERFORMED: yes/no: n/a

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations: 0.0076, 0.076, 0.76, 7.6 were used as range finding concentrations
- Results used to determine the conditions for the definitive study: No effects at these concentrations

Nominal and measured concentrations:

Nominal: 0.0076, 0.076, 0.76, 7.6 (as range finding concentrations) and 76 mg/kg as maximum carbon saturation level.
Geometric mean concentration: (only tested for the nominal 76 mg/kg concentration) 41 mg/kg

Reference substance (positive control):

yes

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 76 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: survival

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 41 mg/kg soil dw

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: survival

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

76 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: survival

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

41 mg/kg soil dw

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: survival

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

> 76 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: weight change

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

> 41 mg/kg soil dw

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: weight change

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

76 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: weight change

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

41 mg/kg soil dw

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: weight change

Duration:

56 d

Dose descriptor:

EC50

Effect conc.:

> 76 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

56 d

Dose descriptor:

EC50

Effect conc.:

> 41 mg/kg soil dw

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

76 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

41 mg/kg soil dw

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: The EC50 value for reproduction was 1.4 mg/kg

Reported statistics and error estimates:

At the termination of each phase of the study (Day 28 for F0 and Day 56 for F1) data obtained on survival and weight change of F0 adults and the number of F1 offspring were statistically analyzed to identify significant treatment-related effects. CETIS-Comprehensive Environmental Toxicity Information SystemTM (Ives, 2013) Version 1.8 software was used to perform the computations.

The No-Observed-Effect Concentration (NOEC), the highest test concentration which demonstrated no statistically adverse effect (p 0.05) when compared to the control data, was determined. The data were first checked for normality using Shapiro-Wilks’ Test (U.S. EPA, 2002) and for homogeneity of variance using Variance Ratio F t Two-Sample Test (U.S. EPA, 2002). If the data sets passed the test for homogeneity and normality, then Equal Variance t Two-Sample Test (U.S. EPA, 2002), was used to determine the NOEC. If the data did not pass the tests for homogeneity and normality, then Unequal Variance t Two Sample Test (U.S. EPA, 2002) was used to determine the NOEC. All statistical conclusions were made at the 95% level of certainty except in the case of the basic assumption tests, e.g., Shapiro-Wilks’ Test and Variance Ratio F Test, in which the 99% level of certainty was applied.

Validity criteria fulfilled:

yes

Conclusions:

28-d F0 LC50 values of >76 mg/kg nominal concentration, >41 mg/kg mean measured concentration, and NOECs of 76 mg/kg nominal concentration, and 41 mg/kg mean measured concentration, have been determined for effects of D5 on survival and weight change of adult earthworms.
56-d F1 LC50 values of >76 mg/kg nominal concentration, >41 mg/kg mean measured concentration, and NOECs of 76 mg/kg nominal concentration, and 41 mg/kg mean measured concentration, have been determined for effects of D5 on reproduction of the earthworm Eisenia fetida.

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Guideline study but it is not certain whether the study was conducted in accordance with GLP.

Qualifier:

according to guideline

Guideline:

other: Environment Canada, EPS 1/RM/43, June 2004

GLP compliance:

not specified

Remarks:

The data is from a publication and this information was not included.

Analytical monitoring:

yes

Details on sampling:

Soil samples were collected throughout the initial treatment, preparation (i.e., at manure amendment, following the 2-h equilibrium period, and following mixing of the amended biosolid with the Gr10 soil) for the range-finding tests to confirm the analytical methodology used, as well as to evaluate the loss of D5 over time. For the definitive tests, samples were taken for analysis only on day 0 of the test (i.e., when the test organisms were added) to confirm dose concentrations in the test soil. All samples were immediately frozen and stored at -20°C prior to analysis. All samples were analyzed for cVMS D4, D5 and D6 concurrently by Quebec Laboratory for Environmental Testing of Environment Canada (Montreal, Quebec).

Triplicate 1.0g (+/-0.2) (wet weight) samples were then measured into glass tubes and well mixed to ensure solids are homogenized. To extract the soil samples, 5 mL of acetonitrile was added to each sample and vortexed followed by the addition of 5 mL of pentane (also vortexed). The sample was then sonicated in an ice bath for 10 min followed by centrifugation at 1500 RPM for 10 min at 15°C. The pentane phase was then transferred to a clean GC vial and 75 μL of naphthalene-D8 solution (100 ng/μL in pentane) was added as an internal standard.

Vehicle:

no

Details on preparation and application of test substrate:

A sandy loam soil (Gr10) was obtained from Greely Sand and Gravel (Ottawa Ontario (ON)) in June 2010 and was dried, sieved (4-mm mesh) and homogenized prior to use. The negative control treatment consisted of a formulated artificial soil (AS) comprising 10% air-dried pre-sieved Sphagnum sp. peat, 20% kaolin clay, and 70% silica sand, with calcium carbonate added to standardize the soil pH.

For the toxicity tests, the soil was amended with either an uncontaminated or contaminated surrogate biosolid. Although available, the use of an actual biosolid produced from a water treatment facility was not pursued as an option for use in the toxicity tests as the potential for other contaminants or toxicants present in the sample may have confounded toxicity test results. As a result, mushroom compost and organic cow manure were both considered as options for a surrogate biosolid. Both alternative biosolids were analyzed in conjunction with a real biosolid (obtained from the Robert O. Picard Water Treatment facility in Ottawa, ON) for comparison of their physical and chemical characteristics. Based on these characteristics. the organic cow manure was selected for its similarity in percent organic matter to the actual biosolid.

To prepare test samples, the surrogate biosolid was amended with D5 (97% pure liquid; Sigma-Aldrich) to create a contaminated test substance; subsequently, the amended biosolid was manually mixed for 2 min, covered to prevent volatilization, and allowed to equilibrate for 2 h. Following the 2 h equilibration period, the amended biosolid was added to the Gr10 soil at a rate of 5 g/kg (on a dry weight basis), equivalent to 8 t/ha, a common application rate for biosolids on agricultural soils in Ontario to create the test soil. Each test concentration was prepared independently. The average moisture content of the test soil was 38 ± 3%, with an average pH of 7.55±0.15 for all test set-ups.

Test organisms (species):

other: Eisenia andrei

Animal group:

annelids

Details on test organisms:

Two sexually mature adults were added to each test vessel at the start of the test (day 0), and the average initial wet weight of the adult earthworm
was 451.7 +/-108.2 mg.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Test temperature:

20 +/-2°C

pH:

7.55 +/-0.15

Details on test conditions:

Test vessels were incubated at 20+/-2°C with an average light intensity of 10.7 +/-2.5 μmol/(m2 t)-1. within the ranges specified in the earthworm method; and food (cooked oatmeal) was added on days 0,14,28 and 42 of the test. On day 28, surviving adults were counted and removed from the test soil, and on day 56. the number of juveniles produced was counted, and the juvenile dry biomass measured and recorded.

Nominal and measured concentrations:

Nominal/measured concentrations (mg/kg dry wt.) of D5 for this test were 0/0, 143/50, 244/91, 414/135, 704/248, 1197/358, 2035/507, 3460/1093, 5882/2362 and 10000/4074.

Reference substance (positive control):

not specified

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 4 074 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

56 d

Dose descriptor:

other: IC50

Effect conc.:

> 4 074 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

56 d

Dose descriptor:

other: IC50

Effect conc.:

> 4 074 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: individual juvenile dry mass

Details on results:

There was no effect on adult survival after 28 d of exposure (100% survival in all treatments, data not shown). While there was a significant reduction in juvenile production at test concentrations ≥507 mg/kg (Fisher's LSD; p<0.05), the lC50 was >4074 mg/kg. There was also no effect on the mean individual dry mass of juveniles; however, the mass increased with increasing exposure to D5. This may be due to less competition for resources (i.e., food) within the test vessel as fewer juveniles were observed at higher test concentrations. Test concentrations > 1093 mg/kg had significantly higher mean individual dry mass compared with lower test concentrations (Fisher's LSD: p < 0.05).

Reported statistics and error estimates:

The lethal concentration (i.e., LC50s) was calculated for invertebrate mortality using the EPA Probit Analysis Program {version 1.5) or the EPA Trimmed Spearman-Karber (TSK) Program (version 1.5). Inhibitory concentrations (i.e., lC50s) were estimated for invertebrate juvenile production using nonlinear regression analysis via SYSTAT (version 13) software, ensuring that model assumptions (normality and homoscedasticity) were met. When regression models were unable to calculate the IC50, or model assumptions were not met, a "Linear Interpolation for Sublethal Toxicity: The Inhibition Concentration (ICp) Approach" (ICPIN) (version 2.0) was used. ln cases where a 50% effect was not observed, an ANOVA was performed to determine if there was a significant difference in the measured endpoint across treatments. lf a significant difference was found. then Fishers least significant difference test was used to determine where the difference was among treatments.

Validity criteria fulfilled:

yes

Conclusions:

A 28-day LC50 value of >4074 mg/kg dry weight and a 56-day NOEC of ≥4074 mg/kg dry weight have been determined for the effects of the test substance on mortality and reproduction and growth respectively of Eisenia andrei.

Description of key information

L4 and L5: LC₅₀ (28-day): >4074 mg/kg dry weight (measured initial concentrations), Eisenia andrei, read-across from D5 (CAS 541-02-6).

NOEC (56-day): ≥4074 mg/kg dry weight (measured initial concentrations), reproduction and growth, Eisenia andrei, read-across from D5 (CAS 541-02-6).

LC₅₀ (28-day): >76 mg/kg (nominal); >41 mg/kg (mean measured), for effects on survival and weight change of adult earthworms, Eisenia fetida, read-across from D5 (CAS 541-02-6). 

NOEC (28-day): 76 mg/kg (nominal); 41 mg/kg (mean measured), for effects on survival and weight change of adult earthworms, Eisenia fetida, read-across from D5 (CAS 541-02-6).

EC₅₀ (56-day): >76 mg/kg (nominal); >41 mg/kg (mean measured) for effects on reproduction of Eisenia fetida, read-across from D5 (CAS 541-02-6).

NOEC (56-day): 76 mg/kg (nominal); 41 mg/kg (mean measured) for effects on reproduction of Eisenia fetida, read-across from D5 (CAS 541-02-6).

Additional information

Constituent HMDS

A stability test under OECD TG 222 conditions with the related substance L3 demonstrated significant loss of test item from the test system. HMDS is more volatile and also more rapidly degradable than L3, meaning that similar losses from the test system would be expected under equivalent test conditions. Additionally, in an OECD TG 216 (Soil Microorganisms: Nitrogen Transformation Test) for the effects of HMDS on nitrate formation rate of soil microflora, analysis of the test substance concentrations show that test material was lost by day three of the test. Based on these experimental findings, the registrants believe it is not technically feasible to proceed with the OECD TG 222 test for HMDS on the basis that the test substance is too volatile to maintain adequate concentrations in the test system.

 

Constituent L3

A stability/recovery test was conducted with L3 in preparation for terrestrial ecotoxicology studies. The study demonstrated a method of introducing neat 14C-octamethyltrisiloxane (14C-L3) into natural soil with subsequent mixing to distribute the test article throughout the soil uniformly. 

The second phase of the study investigated the stability of 14C-L3 in the same soil under conditions representative of those used for the OECD 222 Earthworm Acute Toxicity and Reproduction Test. The system was partially open to allow for respiration during a planned future toxicity experiment. 

Effect of headspace on loss of test material during dosing of soil was examined. The main experiment test vessel was selected in order to minimise headspace as much as possible, while still allowing for enough tumbling to ensure homogeneity. However, even the container with the best recovery only observed an average recovery of 69.7% (or 22.9 mg/kg soil (dw)).

Measures were taken to avoid loss of test substance through volatilisation during the homogeneity experiment. These included:

- fitting the test vessel jar with a polytetrafluoroethylene (PTFE) lined lid and covering the threads of the jar with PTFE tape;

- spiking the soil with a calibrated gastight syringe;

- spiking with a concentration 34% above the saturation concentration, to attempt to account for test substance losses;

- taping the lid closed to avoid inadvertent opening.

The stability experiment was then carried out using the spiked soil from the homogeneity experiment, which was divided into five beakers. Each beaker had 10 mL of headspace and was covered with plastic film having five small holes (approximately 3 mm in diameter) to allow for air exchange. The plastic film was secured with rubber bands. 

By day 3 in the stability experiment, only 6.3% of the initially observed radioactivity was detected, and sampling was stopped. However, the average of the homogeneity determination was taken as the initial concentration for each beaker in the stability test. The soil was not analysed again after dividing into the beakers, therefore the potential loss from moving the test soil from the container used in the homogeneity experiment to the 5 test beakers used in the stability study, was not determined.

A single peak was observed consistent with 14C-L3, showing that the primary mechanism of loss in the initial recovery experiment was volatilisation.

The absence of degradation products in the vast majority of samples, coupled with the rapid loss of 14C activity, shows that the primary mechanism of test article loss was volatilisation of 14C-L3 from the simulated OECD 222 test setup.

Based on these experimental findings, the registrants believe it is not technically feasible to proceed with the definitive OECD 222 test on the basis that the test substance is too volatile to maintain adequate concentrations in the test system.

 

Constituents L4 and L5

There are no data describing the long-term toxicity of L4 and L5 to soil macroorganisms. However, data are available for the siloxane decamethylcyclopentasiloxane (D5, CAS: 541-02-6). A 28-day LC₅₀ value of >4074 mg/kg dry weight and a 56-day NOEC of ≥4074 mg/kg dry weight (measured initial concentration) have been determined for the effects of D5 on mortality and reproduction and growth respectively ofEisenia andrei (Velicogna et al., 2011).

In a second toxicity to soil macroorganisms except arthropods test with D5, no effects were observed at a measured concentration of 41 mg/kg dw. 28-day LC₅₀ values of >76 mg/kg nominal concentration and >41 mg/kg mean measured concentration, and NOECs of 76 mg/kg nominal concentration and 41 mg/kg mean measured concentration, were determined for effects of D5 on survival and weight change of adult earthworms, Eisenia fetida.

In the same test, 56-day EC₅₀ values of >76 mg/kg nominal concentrations and >41 mg/kg mean measured concentration, and NOECs of 76 mg/kg nominal concentration and 41 mg/kg mean measured concentration, were determined for effects of D5 on reproduction of Eisenia fetida (Smithers Viscient, 2015).

 

Testing proposal for Constituent L4

Earthworm reproduction studies are under way for L4. A preliminary stability study under OECD 222 conditions has been carried out and demonstrates significant losses due to volatilisation over a five week period (37% remaining radioactivity after 35 days). However, it is considered that it is possible that measurable concentrations will remain in the soil at the end of the eight-week test period for the definitive OECD 222 study, and therefore testing is proposed.

Read-across of the terrestrial toxicity data for D5 to L4 is considered to be suitable to derive an interim hazard and risk assessment under REACH for L4.

 

 

Testing proposal for Constituent L5

Earthworm reproduction studies are under way for L5. A preliminary stability study under OECD 222 conditions has been carried out using the structurally related substance L4 and demonstrates significant losses due to volatilisation over a five-week period (37% remaining radioactivity after 35 days). However, L5 is expected to be more stable in soil, therefore it is considered that it is possible that measurable concentrations will remain in the soil at the end of the eight-week test period for the definitive OECD 222 study, and testing is proposed.

Read-across of the terrestrial toxicity data for D5 to L5 is considered to be suitable to derive an interim hazard and risk assessment under REACH for L5.