Hydrogen tetrakis[2,2-bis[(allyloxy)methyl]butan-1-olato-O1]bis(ditridecyl phosphito -O'')titanate(2-)

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:

13 - 15 Feb 2019

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

limited significance of analytical dose verification

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen tetrakis[2,2-bis[(allyloxy)methyl]butan-1-olato-O1]bis(ditridecyl phosphito -O'')titanate(2-)

EC Number:

264-709-9

EC Name:

Hydrogen tetrakis[2,2-bis[(allyloxy)methyl]butan-1-olato-O1]bis(ditridecyl phosphito -O'')titanate(2-)

Cas Number:

64157-14-8

Molecular formula:

not applicable (UVCB substance)

IUPAC Name:

dihydrogen 2-{[(di{[bis(tridecyloxy)phosphanyl]oxido}tris({2,2-bis[(prop-2-en-1-yloxy)methyl]butyl}oxido)titaniotetrakis(ylium))oxido]methyl}-1-(prop-2-en-1-yloxy)-2-[(prop-2-en-1-yloxy)methyl]butane

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Remarks:

LC-MS/MS (for reference substance Trimethylolpropane diallyl ether), ICPMS (for reference substance Titanium )

Details on sampling:

- Concentrations (TMPDAE): control, level 1 (nominal loading rate: 0.65 mg/L) , and level 5 (nominal loading rate: 10 mg/L) test solution at initiation (0 hour) and in control and level 1 at termination (48 hours) of the definitive test.
- Concentrations (titanium) : control, level 1 and level 5 test solution samples at initiation (0 hour) of the definitive test. The 0-hour samples were measured in fresh parent test solutions and the 48-hour samples were measured from a composite of the spent.
- Sampling method (TMPDAE): 5-mL sample was collected from the control and levels 1 and 5 test substance treatments for TMPDAE analysis using a serological pipette and placed in separate 10-mL culture tubes.
- Sampling method (titanium): At the 0-hour sampling point, a 5-mL sample was collected from the control and levels 1 and 5 test substance treatments for titanium analysis using a serological pipette, placed in separate 15-mL Falcon™ tubes.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The five test solutions were prepared separately as water accommodated fractions (WAF). Test substance was weighed onto glass cover slips and dropped into glass aspirator bottles containing 2.0-L of dilution water to prepare the test substance solutions. The aspirator bottles were stirred for approximately 22 hours with a vortex <25% of the solution depth. After approximately 22 hours, the solutions were removed from the stir plates and allowed to settle for approximately 1-2 hours. After settling, the level 1 through level 4 (nominal loading rates: 0.65 - 5 mg/L) solutions appeared clear and colorless. The level 5 solution (nominal loading rate: 10 mg/L) appeared clear and colorless with a very light scattered surface film. After the 1-2 hour settling, approximately 100 mL of each solution was discarded, and approximately 1 L of each solution was collected and used to prepare the replicate test treatment chamber solutions.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: obtained from an in-house daphnid culture
- Parental stock: Less than 24 hours old neonates for the definitive test were collected from in-house culture containing adults that were approximately 16 days old. The adults were considered acceptable with no signs of stress, disease or physical damage.
- Feeding during test: no
- Holding conditions: All daphnids were cultured in a temperature-controlled area at approximately 20°C. During the culture period, the daphnids were fed a suspension of the algal species Pseudokirchneriella subcapitata at least once a day supplemented by an artificial diet consisting of a wheat grass, salmon starter, and yeast suspension. Less than 24 hours old neonates for the definitive test were collected from in-house culture containing adults that were approximately 16 days old. The adults were considered acceptable with no signs of stress, disease or physical damage.

ACCLIMATION
- Since the culturing and testing environmental parameters were equivalent (i.e., temperature, dilution water, and lighting), no acclimation period was necessary.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

146 mg CaCO3/L

Test temperature:

20.0°C - 20.6°C

pH:

8.5 - 8.7

Dissolved oxygen:

7.6 - 8.6 mg/L

Conductivity:

335 µS/cm

Nominal and measured concentrations:

Nominal loading rates: 0 (control), 0.65, 1.3, 2.5, 5.0, and 10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 250-mL glass jars
- Type: closed with a clear plastic Petri dish
- Fill volume: 200 mL
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was a moderately hard freshwater prepared by blending naturally hard well water with well water that was demineralized by reverse osmosis (RO). The well water and RO water were blended together to yield a total hardness of 130 to 160 mg CaCO3/L. Prior to use, the dilution water was passed through a sediment filter.

- Culture medium different from test medium: no
- Intervals of water quality measurement: Total hardness, total alkalinity, and conductivity were measured in the dilution water at test initiation. Temperature, dissolved oxygen concentration, and pH were measured at initiation and termination in all replicates.

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: 728 lux

EFFECT PARAMETERS MEASURED: Daphnids were observed for immobility and sublethal effects at approximately 24 and 48 hours after test initiation.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

3.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95%-CI: 2.9 - 4.4 mg/L

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

6.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95%-CI: 6.4 - 7.3 mg/L

Effect concentrations 3

Duration:

48 h

Dose descriptor:

other: NOEL

Effect conc.:

0.65 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: At 24 hours two daphnids in the 1.3 mg/L and two daphnids in the 5.0 mg/L test substance treatments were noted as floating. At 48 hours five daphnids in the 1.3 mg/L and three daphnids in the 5.0 mg/L test
substance treatments were noted as floating

Any other information on results incl. tables

BIOLOGICAL RESULTS:

Table 1: Immobility and percent unaffected of Daphnia magna exposed to KR®55 for 48 hours under static test conditions

 

Nominal Test Substance Loading Rate (mg/L)	Cumulative Immobile (% Immobility)								48-Hour Mean Percentage Immobility	48-Hour Mean Percentage Unaffected a
	24 Hours				48 Hours
	A	B	C	D	A	B	C	D
0 (Control)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0	100
0.65	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0	100
1.3	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0	75 *
2.5	0 (0)	0 (0)	0 (0)	1 (20)	4 (80)	5 (100)	3 (60)	4 (80)	80	20 *
5.0	0 (0)	0 (0)	0 (0)	0 (0)	1 (20)	1 (20)	1 (20)	1 (20)	20	65 *
10	5 (100)	5 (100)	5 (100)	5 (100)	5 (100)	5 (100)	5 (100)	5 (100)	100	0 *

a Percentage unaffected was percentage of daphnids showing no sublethal observations and no immobility at the end of the definitive test (48 hours), with 100% meaning no affected daphnid and 0% meaning all daphnids showing sublethal observations or immobility.

* Statistically significant decrease in percent unaffected compared to the control (Dunnett’s One-Tailed Test and Fisher’s One- Tailed Exact Test with Hochberg’s familywise adjustment).

Notes: Five daphnids were placed in each replicate at test initiation, totaling 20 daphnids per treatment.

ANLYTICAL RESULTS:

TMPDAE:

Table 2: Measured concentrations of TMPDAE during the 48-hour acute toxicity test with Daphnia magna to KR®55

Nominal KR®55 Test Substance Loading Rate (mg/L)	Measured TMPDAE Concentrationaas mg/L
	0 Hour	48 Hours	Mean Measured
0 (control)	<LOD b	<LOD b	<LOD b
0.65	0.139	0.133	0.136
1.3c	NA	NA	NA
2.5c	NA	NA	NA
5.0c	NA	NA	NA
10	2.27	--e	NA
QC Fortification Spikes (% Recovery)
Low Spike (0.00194) d	0.00205 (105) 0.00199 (103) 0.00195 (100)	0.00200 (103) 0.00206 (106)	NA
High Spike (121) d	121 (100) 127 (105) 125 (104)	121 (100) 127 (105)	NA

a Measured Concentration (mg/L) = Measured concentration from curve (ng/mL) × analysis volume (mL) / sample volume (mL) / 1,000 (ng/L / mg/mL).

b LOD = 0.000581 mg/L.

c TMPDAE concentrations measured in lowest and highest treatments only. No samples collected from levels 2, 3, and 4.

d Nominal concentration in mg TMPDAE/L.

e No sample collected due to 100% immobility.

NA = Not applicable.

Titanium:

Table 3: Measured concentrations of titanium at 0 Hour During the 48-Hour acute toxicity test with Daphnia magna to KR®55

 

Nominal KR®55 Test Substance Loading Rate (mg/L)	Measured Titanium Concentrationaas mg/L
	0 Hour
0 (control)	<LODb
0.65	<LODb
1.3 c	NA
2.5 c	NA
5.0 c	NA
10	<LODb
QC Fortification Spikes (% Recovery)
Low Spike (0.0100)d	0.00753 (75)
Mid Spike (0.100) d	0.0992 (99)e
High Spike (10.0) d	10.3 (103)f

aMeasured Concentration (mg/L) = Measured concentration from curve (mg/L) × analysis volume (mL) / sample volume (mL).

b LOD = 0.00300 mg/L.

cTitanium concentrations measured in lowest and highest treatments only. No samples collected from levels 2, 3, and 4.

d Nominal concentration in mg titanium/L.

e Re-diluted in duplicate and re-analyzed, mean of duplicate re-analyses reported.

f Re-diluted in duplicate and re-analyzed, mean of original and duplicate re-analyses reported.

NA = Not applicable.

VALIDITY CRITERIA:

Table 4: Validity criteria for OECD 202.

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	Immobilization among control daphnids was 0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	The dissolved oxygen concentration at the end of the test was ≥ 8.3 mg/L in control and test substance treatments	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.