Tributyl-2-methoxypropylphosphonium salt with 4,4'-[2,2,2-trifluoro-1-(trifluoromethyl)ethylidene]bis[phenol]

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 July 2011 - 24 Aug 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study under GLP with analytical determination of concentrations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

MTDID 3285

IUPAC Name:

MTDID 3285

Details on test material:

- Name of test material (as cited in study report): MTDID 3285, CAS# 126049-00-1
- Substance type: Technical grade substance
- Physical state: Pink waxy solid with lumps
- Analytical purity: 86.67%
- Lot/batch no.: DZ-2200 - lot 30069
- Expiration date of the lot/batch: 31 May 2014
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: WSFs prepared at loading rates of 1.0, 10 and 100 mg/L for the rangefinding test, and 1.0, 2.2, 4.6 and 10 mg/L for the definitive test. A 46% dilution of the 1.0 mg/L loading rate was also prepared for the definitive test.

- Sampling method: Aliquots (2 mL) were taken from all test concentrations and the control at times 0 and 48 hours.

At the end of the test, replicates were pooled at each concentration before sampling.

- Sample storage conditions before analysis: Samples were stored frozen (≤-15 °C) until the day before analysis, thawed, diluted with an equal volume of methanol, and held overnight at room temperature before analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water soluble fraction. Loading rates of 1.0, 2.2, 4.6 and 10 mg/L were sonicated for ten minutes and then were stirred for three days. Undissolved material was removed by filtration through 0.45 µm membranes. Portions of the solution loaded at 1.0 mg/L was then further diluted to produce a 46% dilution. The final test solutions were all clear and colourless.
- Controls: Blank
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None after the filtration step

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Daphnia magna Straus, 1820
- Source: In-house laboratory culture
- Age at study initiation (mean and range, SD): < 24 hours
- Method of breeding: Acyclical parthenogenesis. Breeding batch started by placing approximately 250 daphnids less than 3 days old into 5 liters of M7 medium (modified ISO medium) in an all-glass culture vessel. Cultures maintained for a maximum age of 4 weeks. After the first 7 days, cultures are renewed by replacement of half of the medium twice weekly. Cultures are maintained at 18 - 22 deg. C and fed daily with a suspension of freshwater algae.
- Feeding during test: None

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg CaCO3 / L

Test temperature:

19.7 °C - 20.8 °C

pH:

7.9-8.0

Dissolved oxygen:

8.8 - 9.1 mg/l

Salinity:

NA

Nominal and measured concentrations:

Nominal concentrations: Based on WSF loading rates: Control (blank), 0.46* mg/L loading rate, 1.0 mg/L, 2.2 mg/L, 4.6 mg/L and 10 mg/L
*(0.46 mg/L solution was a 46% dilution of the 1.0 loading rate)

Mean measured concentrations (based on anion): <0.002 mg/L, 0.34 mg/L, 0.81 mg/L, 1.3 mg/L, 3.6 mg/L, 7.4 mg/L (initial concentrations. See Table 1)

Details on test conditions:

TEST SYSTEM
- Test vessel: 100-mL all-glass containers. Fill volume was 80 mL.
- Aeration: None
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): four
- No. of vessels per control (replicates): four

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water used to make growth medium was tap water purified by reverse osmosis.
- Culture medium different from test medium: no, standard M7 medium used throughout. M7 is based on standard ISO medium supplemented with additional minerals.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h:8 h light:dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Immobilization: Daily
-Mortality: Daily

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 2.2 (1/3 log unit)
- Range finding study: Yes- Test concentrations: WSFs based on 1.0 mg/L, 10 mg/L and 100 mg/L loading rates, plus 10% dilution of 1.0 mg/L WSF
- Results used to determine the conditions for the definitive study: Yes. EC50 between 1.0 and 10 mg/L loading rate.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, reference test done 2 weeks after the definitive MTDID 3285 test

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

4.1 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95% CI, 3.5-5.0 mg/L

Details on results:

Measured concentrations at beginning and conclusion of test are shown in Table 1. Concentrations were essentially identical at conclusion of the test as at the beginning, therefore initial concentrations were used to calculate EC50. Because anion concentrations were somewhat lower than concentrations of the cation, EC50 is calculated using anion measurements.

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No. Test solutions were clear and colorless after filtration. There was no indication that the standards contained any insoluble material

- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

In a test of potassium dichromate conducted 2 weeks after the definitive test, a 48-h EC50 of 0.41 mg/L was observed. This EC50 is within the range of values for reference tests of D. magna performed by the testing lab.

Reported statistics and error estimates:

Immobilization data are in Table 2. Immobilization of 5% at 0.81 was not considered biologically significant

EC50 was calculated at 48 hours of exposure from the probits of the percentages of
affected daphnids and the logarithms of the corresponding test substance concentrations using the maximum likelihood estimation method.

Any other information on results incl. tables

Table 1, Concentrations of MTDID 3285 in the test medium
Day of sampling (age)	Concentration
	Loading rate (mg/I)	Analysed(mg/l)	Percent of initial concentration
0 hours	0	<0.002	─
	46% of 1.0	0.341	─
	1.0	0.805	─
	2.2	1.29	─
	4.6	3.60	─
	10	7.44	─
48 hours	0	<0.002	─
	46% of 1.0	0.313	92
	1.0	0.770	96
	2.2	1.47	114
	4.6	3.64	101
	10	8.01	108

Table 2, Acuteimmobilisationof daphnia after 24 and 48 hours in the definitive test
MTDID 3285 initial concentration (mg/L)	Vessel number	Number Daphnia exposed	Response at 24 h		Response at 48 h
			Number	% of Total	Number	% of Total
Control (<0.002)	A	5	0	0	0	0
	B	5	0		0
	C	5	0		0
	D	5	0		0
0.34	A	5	0	0	0	0
	B	5	0		0
	C	5	0		0
	D	5	0		0
0.81	A	5	0	0	1	5
	B	5	0		0
	C	5	0		0
	D	5	0		0
1.3	A	5	0	0	0	0
	B	5	0		0
	C	5	0		0
	D	5	0		0
3.6	A	5	2	15	3	40
	B	5	1		2
	C	5	0		2
	D	5	0		1
7.4	A	5	3	60	5	100
	B	5	4		5
	C	5	2		5
	D	5	3		5

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

<10% immobilization in controls, dissolved oxygen >3 mg/L

Conclusions:

The 48-hour EC50 of MTDID 3285 to Daphnia magna was 4.1 mg/L, 95% CI 3.5 - 5.0 mg/L (OECD TG202)

Executive summary:

The toxicity of MTDID 3285 to Daphnia magna (water flea) was assessed in a static test conducted according to OECD TG202. Water-soluble fractions (WSFs) were made using loading rates of 1.0, 2.2, 4.6 and 10 mg/L, and the 1.0 mg/L loading rate was additionally diluted to 46% to cover a lower concentration range. Analytically measured concentrations were essentially identical at beginning and conclusion of the test, so the measured initial concentrations of the anion were used to calculate EC50. Since recovery was greater for the cationic portion of the substance relative to the anionic portion, the EC50 calculation based on the anion provides more conservative result and was reported here. The 48-hour EC50 of MTDID 3285 to Daphnia magna was 4.1 mg/L based on immobilization.

The study was conducted under accepted guidelines and according to GLP criteria. The test material contained an unspecified amount of insoluble material which was removed by filtration. There was no indication that the standards, which were made in 50% (v/v) methanol, contained any undissolved material. Also, while analytical results for both chemicals were constant for both portions of the molecule, measured anion concentrations were somewhat lower at test initiation than cation concentrations. Therefore, an EC50 based on anion concentrations is more conservative than an EC50 based on cation concentrations. The study is classified as reliable and is suitable for use in classification and labelling.

Results Synopsis

Test Type: Static

EC50: 4.1 mg/L                         95% C.I.:  3.5 - 5.0 mg/L

NOEC: NA                 Probit Slope:  log 10 (conc) = 0.12 + (probit - 2.84)/4.43