Amino functional alkoxysilanes

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

19th of September 2019 to 15th of January 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1300 (Daphnid Chronic Toxicity Test)

Version / remarks:

1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2012

Deviations:

yes

Remarks:

Only one concentration used (rather than five)

GLP compliance:

yes (incl. QA statement)

Remarks:

In total, three exceptions are listed, however, these are not expected to have provided any impact on the quality of the study.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (water control) and 12 mg methylsilanetriol/L
- Sampling method: The bulk water control and bulk test solution were sampled at test initiation (day 0) and on days 2, 7, 14 and 19 of the exposure. Spent (pooled replicate) solutions were sampled on days 2, 5, 7 and 14 as well as at exposure termination (day 21). To assess analytical method precision and solution homogeneity, three additional samples were collected on day 0, 7, 14 and 19 from the bulk test solution. The collected samples were analysed for methylsilanetriol by HPLC/MS-MS. Validation of the analytical methodology was conducted concurrently with the study as per standard operating procedure (SOP) MTH-ACL-2 (2014).
- Sample storage conditions before analysis: Not reported.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The definitive test was conducted under static-renewal conditions for 21 days. Ten replicate test vessels, each containing test solution or LDW were prepared per test level. At test initiation, one daphnid was impartially added to each test vessel within 30 minutes of the addition of the test solutions. The replicate test vessels were placed into a temperature-controlled environmental chamber according to a computer-generated randomization.
- Eluate: not specified
- Differential loading: not specified
- Controls: water
- Chemical name of vehicle: n/a
- Concentration of vehicle in test medium: n/a
- Evidence of undissolved material: not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: n/a
- Justification for species other than prescribed by test guideline: n/a
- Age at study initiation: Less than 24 hours
- Weight at study initiation: n/a:
- Length at study initiation: Mean length per surviving adult female were 4.81 mm in the water control and 4.85 mm in the 10.0 mg/L test level.
- Stage and instar at study initiation: n/a
- Valve height at study initiation, for shell deposition study: n/a
- Peripheral shell growth removed prior to test initiation: n/a
- Method of breeding: n/a
- Source: In house cultures obtained originally from Aquatic BioSystems, Inc., Fort Collins, Colorado
- Age of parental stock: n/a
- Feeding during test : Daphnids were fed with Raphidocelis subcapitata and YTC (yeast, wheat grass, finfish food suspension) daily from the test initiation (day 0) to day 20, however, not on day 21.
- Food type: Raphidocelis subcapitata and YTC (yeast, wheat grass, finfish food suspension)
- Amount: 0.8 mL Raphidocelis subcapitata and 0.1 mL YTC (yeast, wheat grass, finfish food suspension) daily between day 0-6. Thereafter, 0.9 ml of Raphidocelis subcapitata and 0.1 mL YTC between day 7-20. No food was provided on day 21. Therefore, the final feeding ration resulted in approximately 0.2 mg C/daphnid/day throughout the entire exposure period.
- Frequency: daily

ACCLIMATION
- Acclimation period: n/a
- Acclimation conditions: n/a
- Type and amount of food: n/a
- Feeding frequency: n/a
- Health during acclimation: not specified

QUARANTINE
- Duration: n/a
- Health/mortality: n/a

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS: n/a

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Post exposure observation period:

n/a

Hardness:

58-62 mg CaCO3/L

Test temperature:

19–20°C

pH:

7.7-8.9

Dissolved oxygen:

8.0–10.5 mg/L (90–118% air saturation)

Salinity:

not specified

Conductivity:

160-170 µS/cm

Nominal and measured concentrations:

Nominal test concentration: 0 (water control) and 12 mg methylsilanetriol/L
Mean measured test concentrations: < LLQ (water control) and 10.0 mg methylsilanetriol/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 125-mL glass borosilicate jars containing approximately 90 mL of test solution. Each test vessel was loosely covered and uniquely labeled for identification purpose (i.e. replicate, study, test level). Test vessels were replaced with new/clean test vessels on renewal days.
- Type: closed
- Material/size/headspace/fill volume: Glass/125 ml/not specified/90 ml
- Aeration: not specified
- Type of flow-through: Septum Purge Flow (for the GC)
- Renewal rate of test solution (frequency/flow rate): 1 mL/min
- No. of organisms per vessel: 1 daphnia per vessel
- No. of vessels per concentration: 10
- No. of vessels per control: not specified
- No. of vessels per vehicle control: n/a
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water was obtained from the upper Saginaw Bay of Lake Huron off Whitestone Point and was limed and flocculated with ferric chloride. Before use in the laboratory, the water was sand-filtered, pH-adjusted with gaseous CO2, carbon-filtered, and UV-irradiated.
- Total organic carbon: 1.9 mg/L,
- Particulate matter: < 2 mg/L
- Metals/pesticides/chlorine/alkalinity/Ca/mg ratio: concentrations of contaminants were below levels of concern that may have interfered with the results of this test.
- Conductivity: 160-170 μS/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: Weekly

OTHER TEST CONDITIONS
- Adjustment of pH: Yes, with gaseous CO2
- Photoperiod: 16 hours light/8 hours dark per day
- Light intensity: Not exceeding 1000-1500 lux.

EFFECT PARAMETERS MEASURED: not specified

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not specified
- Justification for using less concentrations than requested by guideline: not specified
- Range finding study: No, based on available literature
- Test concentrations: 12 mg methylsilanetriol/L
- Results used to determine the conditions for the definitive study:

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Remarks:

MATC

Effect conc.:

> 10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality: Number of living offspring produced per surviving parental animal {for Daphnia magna, TG 211}

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality: Number of living offspring produced per surviving parental animal {for Daphnia magna, TG 211}

Details on results:

- Behavioural abnormalities: No
- Observations on body length and weight: The daphnid lengths in the water control were not statistically different to the mean lenght per surviving adult female. No weight is recorded.
- Other biological observations: All biological data were normally distributed and homogenous.
- Mortality of control: 0 %
- Other adverse effects control: None reported
- Immobilisation of control: None reported
- Abnormal responses: None reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: Yes

Reported statistics and error estimates:

Shapiro-Wilk’s test at a Type I error rate of 0.01 was utilised for the testing of normality for reproduction and growth (length). To evaluate the homogeneity of variance, Levene's test at a type I error rate of 0.05 was used. Furthermore, a parametric analysis (ANOVA) was conducted using a Dunnett's test to compare the limit test level with the water control. one-tailed Dunnett’s test, looking for a significant decrease from the water control, was conducted at a Type I error rate of 0.05. Additionally, the standard deviation, coeffcient of variance and mean was calculated for e.g. the individual observations

Validity criteria fulfilled:

yes

Conclusions:

A 21 day NOEC value of 10 mg/l (mean measured concentration) (highest concentration tested) has been determined for the effects of methylsilanetriol on reproduction, growth (length) and mortality of Daphnia magna. Additionally, LOEC for the same endpoints was determined to be >10 mg/L.

Executive summary:

A 21 day chronic toxicity study in Daphnia magna with the test substance methylsilanetriol (pre-hydrolysed from the parent substance methyltrimethoxysilane) was conducted according to OECD TG 211 and in compliance with GLP. A 21 day NOEC of 10 mg/l (mean measured concentration) (highest concentration tested) was determined for the effects of methysilanetriol on reproduction, growth (length) and mortality. Additionally, a LOEC for the same endpoints was determined to be >10 mg/L.

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not reported

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Guideline study with acceptable restrictions. The study was not conducted under GLP and exposure concentrations were not measured.

Qualifier:

no guideline followed

Principles of method if other than guideline:

The study consisted of triplicate runs of a 21-day test. Each run consisted of duplicates for control and all test concentrations. Each duplicate started wlth five 24-48 hour old Daphnia magna. Concentrations of 0.01, 0.03, 0. 1 , 0.3 and 1.0 ppm were selected for the test. First generation Daphnia mortality was recorded throughout the test and their reproductive functlon was assessed by counting the total number of young produced through day 21.

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

The substance was added directly to 600 mL beakers, each containing 400 ml of hard reconstituted fresh water.

Test organisms (species):

Daphnia magna

Details on test organisms:

- Source: The test daphnlds were from a laboratory culturing. The origlnal colony of Daphnia magna was obtained through the Dow Chemical Company, which obtained thelr colony from the Duluth Laboratory of the Unlted States Envlronmental Protection Agency.

- Culture medium: The colonies were maintained under static conditions in reconstltuted hard water (EPA 660/3-75-009) made-up from glass-distilled water. Before culture inoculation, the reconstituted water was aerated for 2-4 hours.

- Culture conditions: The colonies were maintained in five-gallon glass aquaria under approximately 1000 foot-candle llluminatlon (18/6 hour light/dark) in an environmental chamber designed to maintain 22+/-1°C.

- Feeding: The food for the Daphnia. magna is prepared each week by blending 0.25 g alfalfa meal, 4.0 g trout chow and 400 mL dlstilled water. Thls solution is allowed to settle and the supernatant is used as food.

,- Test organisms; A number of egg-bearing daphnia were isolated prior to each test for a period of 24-48 hours. Five 24-48 hour old
Daphnia magna produced in the period prior to the test were introduced into each test concentration along with the untreated control group.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

No data

Test temperature:

22+/-1°C

pH:

No data

Dissolved oxygen:

No data

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal concentrations: 0 (Control), 0.01, 0.03, 0.1, 0.3 and 1.0 ppm

Details on test conditions:

Five 24-48 hour old Daphnia magna were introduced into each test concentration along with the untreated control group.

The test consisted of tripllcate runs of the 21-day test. Each run consisted of duplicates for control and all test concentrations, each duplicate started wlth five 24-48 hour old Daphnia magna.

The daphnia were fed 0.25 ml of daphnia food each day except weekends throughout the 21-day study.

0bservations were made each day for each of the test concentrations and control groups for mortalities and young produced.

The daphnia were transferred to fresh test and control vessels weekly throughout the 2l-day study.

Survival of the first generation daphnids was recorded throughout the test. The reproductive functlon of the flrst generation daphnia was assessed by counting the total number of young produced through day 21.

Comparlson between test vessels and controls were made for first generation survival, fecundity, and survlval of the second generatlons. General
observation of behavior and abnormalities were also made.

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 other: ppm

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: reproduction and mortality

Details on results:

Control parental daphnia survival after 21 days: 96.7%

There were no statistically significant differences in survival and reproductive output between the Controls and any of the treatments.

Reported statistics and error estimates:

The levels of effect on survival and reproduction were insufficient to determine ECx values.

Differences in reproductive output were assessed by t test with p<0.05

Table 1. Test results

 

Nominal concentration (ppm)	Survival of parental daphnia (%)	Total young produced +/-Stand Dev	Young produced per female +/-Stand Dev	Initial survival of offspring (%)
0 (Control)	96.7	52.7+/-25.3	10.72+/-4.81	99.7
0.01	100	47.7+/-23.3	9.53+/-4.66	98.3
0.03	100	46.0+/-28.1	9.20+/-5.63	100
0.1	100	52.2+/-25.3	10.43+/-5.07	99.7
0.3	100	44.2+/-29.0	8.83+/-5.81	99.6
1.0	90	33.5+/-17.1	6.93+/-3.19	99.5

 

Validity criteria fulfilled:

yes

Conclusions:

A 21-day NOEC of ≥1 ppm has been determined for the effects of the test substance on survival and reproduction of Daphnia magna. The results are expressed relative to nominal concentrations of the test substance. However the substance is subject to rapid hydrolysis and under the test conditions it is therefore likely that exposure will have been to its hydrolysis products (methanol and N-(3-(trihydroxysilyl)propyl)ethylenediamine).

Description of key information

Long-term toxicity to invertebrates:

Silanol HP-X - read-across from methylsilanetriol (CAS 2554-53-6): 21-day NOEC ≥10 mg/l, mortality, reproduction and growth of Daphnia magna (Dow Chemical Company, 2019).

Silanol HP-Z - read-across from N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3): 21-day NOEC >0.8 mg/l, survival and reproduction of Daphnia magna

(Dow Corning, 1980).

Additional information

Although not required to be included in the REACH data set at this annex level, consideration has been given to the potential long-term toxicity of the registration substance.

The majority of the constituents of the substance are expected to hydrolyse rapidly, therefore the environmental aspects of chemical safety assessment are based on the hydrolysis products, which are grouped into four silanol assessment entities Silanol HP-W, Silanol HP-X, Silanol HP-Y and Silanol HP-Z. However, hydrolysis rates of some of the constituents of the substance are uncertain, therefore risk assessment has also been performed on the parent constituents which are divided into separate Blocks. The same data are used to derive PNEC for the parent Block, depending on the hydrolysis product(s) of the Block. This is discussed further in the ecotoxicological general discussion.

 

Long-term aquatic invertebrate toxicity data are available for two of the four silanol assessment entities and are presented below:

Silanol HP-X: A 21-day NOEC of ≥10 mg/l (measured concentrations) was determined for the effects of methylsilanetriol (CAS 2554-53-6) on mortality, reproduction and growth of Daphnia magna. The study was conducted according to a standard guideline (OECD TG 211) and in compliance with GLP (Dow Chemical Company, 2019).

Silanol HP-Z: A 21-day NOEC of ≥1 ppm (≥1 mg/l) has been read-across from a structural analogue, N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3) on survival and reproduction of Daphnia magna (Dow Corning, 1980). The study was conducted according to a methodology similar to that recommended in the standard guidelines for this endpoint, although not in accordance with GLP. It is likely that the test organisms were exposed predominantly to the hydrolysis products of the substance, N-(3-(trihydroxysilyl)propyl)ethylenediamine (1 mole) and methanol (3 moles). Methanol is not likely to have contributed to toxicity at the tested concentrations. The NOEC value expressed in terms of N-(3-(trihydroxysilyl)propyl)ethylenediamine is ≥0.8 mg/l.

No long term data are available for Silanol HP-W and Silanol HP-Y.