Bis[2-[(2-aminoethyl)amino]ethanolato][2-[(2-aminoethyl)amino]ethanolato-O](propan-2-olato)titanate

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:

03 - 05 Apr 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Version / remarks:

(2004)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Bis[2-[(2-aminoethyl)amino]ethanolato][2-[(2-aminoethyl)amino]ethanolato-O](propan-2-olato)titanate

EC Number:

265-723-8

EC Name:

Bis[2-[(2-aminoethyl)amino]ethanolato][2-[(2-aminoethyl)amino]ethanolato-O](propan-2-olato)titanate

Cas Number:

65380-84-9

Molecular formula:

not applicable (UVCB substance)

IUPAC Name:

2-methyl-4,4-bis(propan-2-yloxy)-3,5-dioxa-8-aza-4-titanadecan-10-amine; 7,7-bis(propan-2-yloxy)-6,8-dioxa-3,11-diaza-7-titanatridecane-1,13-diamine; 7,7-bis({2-[(2-aminoethyl)amino]ethoxy})-6,8-dioxa-3,11-diaza-7-titanatridecane-1,13-diamine; 7-{2-[(2-aminoethyl)amino]ethoxy}-7-(propan-2-yloxy)-6,8-dioxa-3,11-diaza-7-titanatridecane-1,13-diamine; tetrakis(propan-2-yloxy)titanium

Sampling and analysis

Analytical monitoring:

yes

Remarks:

ICP-MS

Details on sampling:

- Concentrations: Control, level 1 (6.5 mg/L nominal loading rate), and level 5 (100 mg/L nominal loading rate) test solution samples at initiation (0 hour) and termination (48 hours) of the test. 0-hour samples were measured in fresh parent test solutions and the 48-hour samples were measured from a composite of the spent replicate solutions.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The five test solutions were prepared separately as water accommodated fractions. Test substance was weighed onto polypropylene funnels and dropped into glass aspirator bottles containing 2.0-L of dilution water being stirred on a stir-plate using a 2-inch magnetic stir bar. The aspirator bottles were stirred for approximately 20 hours with a vortex ≤25% of the solution depth. After approximately 20 hours, the stirring was stopped and the solutions allowed to settle for approximately 1 hour. After settling, the control, level 1 (nominal loading rate: 6.5 mg/L), and level 2 (nominal loading rate: 13 mg/L) solutions appeared clear and colorless with no surface film. The level 3 (nominal loading rate: 0.25 mg/L) solution was cloudy with no surface film. After the 1 hour settling, approximately 100 mL of each solution was discarded, and approximately 1 L of each solution was collected and used as the 6.5, 13, 25, 50, and 100 mg/L test treatment solutions.
- Differential loading: yes
- Controls: WAF preparation consisting of dilution water only
- Evidence of undissolved material: The level 4 (nominal loading rates: 50 mg/L) solution was cloudy with a layer of fine white substance settled on the bottom of the bottle. The level 5 solution was cloudy, with a layer of fine white substance on the bottom of the bottle and a light surface film.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: in-house daphnid culture
- Holding conditions: 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.
- Age of parental stock: adults that were approximately 19 days old
- Feeding during test: no

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

Test temperature:

20.1°C - 20.8°C

pH:

8.5 - 8.8

Dissolved oxygen:

7.8 - 8.7 mg/L

Conductivity:

320 µS/cm

Nominal and measured concentrations:

Nominal loading rates: 0 (control), 6.5, 13, 25, 50, and 100 mg/L
Mean measured titanium concentrations: < LOD (control), 0.453 (6.5 mg/L nominal loading rate), 2.99 (100 mg/L nominal loading rate)

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 mL glass jars
- Type: closed with a Petri dish
- Fill volume: 200 mL
- Aeration: no
- 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: 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.
- Total organic carbon: < 2.0 mg/L
- Culture medium different from test medium: no
- Total alkalinity: 148 mg CaCO3/L
- Hardness: 136 mg CaCO3/L
- Intervals of water quality measurement: Total hardness, total alkalinity, and conductivity were measured in the dilution water, as represented by the control WAF solution, at test initiation. Temperature, dissolved oxygen concentration, and pH were measured in the parent treatment solutions at initiation and in all replicates treatment solutions at termination.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: 591 - 643 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):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

89 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Remarks on result:

other: 95% confidence limit: 73 - 122 mg/L

Effect concentrations 2

Duration:

48 h

Dose descriptor:

other: NOEL

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Details on results:

Floating daphnids: one daphnid in the 6.5 mg KR®44/L test substance treatment, three daphnids in the 50 mg KR®44/L test substance treatment, and one daphnid in the 100 mg KR®44/L test substance treatment at 24 hours. At 48 hours, floating was observed in one daphnid in the 25 mg KR®44/L test substance treatment and nine daphnids in the 50 mg KR®44/L test substance treatment. No other sublethal effects were observed.

Reported statistics and error estimates:

All statistical analyses were performed with SAS software (version 9.3). When possible, estimates of EC50 values and their 95% confidence limits were calculated using the probit model or the trimmed Spearman-Karber method. When the p-value for Goodness of Fit was >0.05 and there was no other evidence of questionable convergence, the probit method was selected for reporting. When this criterion was not achieved, the trimmed Spearman-Karber method was selected for reporting. The no-observed-effect concentration (NOEC) was determined by using a Dunnett’s one-tailed test and a Fisher’s one-tailed exact test with Hochberg’s familywise adjustment.

Any other information on results incl. tables

Biological results:

Table 1: Immobility and percent unaffected of Daphnia magna exposed to KR®44 for 48 hours under static test conditions

 

Nominal Test Substance Loading Rate (mg KR®44/L)	Cumulative Immobile (% Immobility)								48-Hour Mean Percentage Immobility	48-Hour Mean Percentage Unaffecteda
	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
6.5	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0	100
13	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0	100
25	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)	0	95
50	0 (0)	0 (0)	0 (0)	0 (0)	1 (20)	0 (0)	1 (20)	0 (0)	10	45 *
100	0 (0)	0 (0)	0 (0)	0 (0)	3 (60)	2 (40)	4 (80)	3 (60)	60	40 *

aPercentage Unaffectedwaspercentageofdaphnids showingnosublethal observations and no immobility at the end of the definitive test (48hours),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.

Analytical results:

Table 2: Measured concentrations of titanium analytical marker during the 48 hour acute toxicity test with Daphnia magna to KR®44

 

Nominal KR®44 Test Substance Loading Rate (mg KR®44/L)	Measured Titanium Concentrationaas mg/L
	0 Hour	48 Hours	Mean Measured
0 (control)	<LODb	<LODb	<LODb
6.5	0.514	0.393	0.453
13c	NA	NA	NA
25c	NA	NA	NA
50c	NA	NA	NA
100	3.19	2.79	2.99
QC Fortification Spikes (% Recovery)
Low Spike (0.0100)d	0.00988 (99) 0.00864 (86)	0.00840 (84) 0.00885 (89)	NA
Mid Spike (0.100)d	0.0936 (94)	0.0900 (90)	NA
High Spike (10.0)d	9.26 (93) 9.18 (92)	9.58 (96) 9.56 (96)	NA

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

b Titanium LOD = 0.00300 mg/L.

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

d Nominal concentration in mg titanium/L.

NA = Not applicable.

Validity criteria:

Table 3: 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.	0%	yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	7.8 - 8.7 mg/L throughout the test	yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

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