Dihydrogen hexahydroxyplatinate, compound with 2-aminoethanol (1:2)

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Samples with five different nominal concentrations of 4.1, 9.1, 20.0, 44.0 and 96.9 mg test item/L were analysed. Sampling took place at the test start, after 24 hours and at the end of the test (48 h). For the 48-h samples old and fresh samples were analysed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a total amount of 3.608 g of the test item was weighted out and dissolved in 4.0 L of dilution water by mixing for approximately 1 hour. After stirring, the stock solution was filtered via a PES-bottle top filter. Additionally, the stock solution served as highest test concentration. The remaining test concentrations were prepared by serial dilution with dilution water. After 48 hours, new test concentratins were prepared following the same approach.
- Controls: dilution water only
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): none

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain/clone: clone V
- Source: German Federal Environment Agency, Institut für Wasser-, Bodenund Lufthygiene
- Age of parental stock: 3 weeks old
- Age of test organisms at test initiation: 4 - 24 hours old
- Feeding during test: none

ACCLIMATION
- Acclimation period: few hours
- Health during acclimation (any mortality observed): not reported

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: adult Daphnia were separated from the stock population (bred in the laboratory of the Fraunhofer IME) by sieving. Batches of 30 to 50 animals were held at room temperature in about 1.8 L dilution water for one week. During this week the daphnids were fed daily with an algal suspension (Desmodesmus subspicatus) and LiquizeIIR (HOBBY). Algae growing in the log-phase were centrifuged and the pellet was resuspended in a few mL of medium. 30 mL of this suspension was given to 1 L Daphnia medium. The water was changed once per week. Newborn D. magna were separated by sieving, the first generation was discarded. Individuals applied in the test were transferred with a bore Pasteur pipette a few hours after sieving to ensure applying only healthy organisms.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

1.1 - 1.2 mmol/L

Test temperature:

20.0 - 20.5 °C

pH:

8.1 - 8.6

Conductivity:

207 - 242 μS/cm

Nominal and measured concentrations:

Nominal concentrations: 38.5, 84.7, 186.4, 410 and 902 mg test item/L (corresponding to mean measured concentrations of 2.3, 5.3, 11.5, 25.3 and 54.8 mg 2AE/L, respectively, and 0.42, 0.94, 2.17, 4.86 and 10.9 mg Pt/L, respectively)

Details on test conditions:

TEST SYSTEM
- Test vessel: 50-mL glass beakers, filled with about 50 mL test solution and covered with glass panes
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): freshly prepared test solutions were used after 24 hours
- 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: purified drinking water used as holding- and dilution water. The purification included filtration with activated charcoal, passage through a limestone column and aeration. To avoid copper contamination, plastic water pipes were used for the testing facilities.
- Total Hardness: 1.1 - 1.2 mmol/L
- Metals: Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn < LOQ
- Chlorine: < 0.02 - 0.02
- Alkalinity: 1.8 mmol/L
- Conductivity: 207 - 242 μS/cm
- Intervals of water quality measurement: oxygen concentration, pH and temperature of the test solutions were checked at test start and end.

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours dark
- Light intensity: 989 - 985 lux (corresponding to 13.1 – 13.2 μE/(m² *s))

EFFECT PARAMETERS MEASURED: the number of dead animals was visually determined daily. Any abnormalities in appearance and behaviour were recorded. Immobility was determined according to the OECD guideline 202. Organisms which were not able to swim within 15 seconds after gentle agitation of the test vessel were considered to be immobilised (even if they were still moving their antennae).

RANGE-FINDING STUDY - none

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 902 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 54.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

other: 2-amino-ethanol (2AE)

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 10.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 902 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 54.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

other: 2-amino-ethanol (2AE)

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 10.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mobility

Details on results:

- Neither significant signs of disease nor stress like discolouration or abnormal behaviour were observed in any replicate up to a nominal test item concentration of 902.0 mg test item/L. All surviving organisms, up to the highest test item concentration, gave the impression of healthy condition.
- The 48-h LOEC was determined to be > 902 mg test item/L (corresponding to > 54.8 mg 2AE/L and > 10.9 mg Pt/L).

Results with reference substance (positive control):

The most recent results obtained for the reference substance (March 2015) were in agreement with historical 24 hours EC50 values obtained in the Fraunhofer IME Institute. a 24-h EC50 of 1.38 mg reference substance/L was obtained based on immobilisation of Daphnia magna.

Reported statistics and error estimates:

For immobilization, the NOEC and LOEC were determined. EC10, EC20, and EC50 values together with 95% confidence intervals (where possible) were calculated by Probit-analysis assuming log-normal distribution of the values. All statistical calculations were performed using the computer programme ToxRat.

Table 1. Mobile daphnids and % immobilisation after 24 and 48 h of exposure to dihydrogen hexahydroxyplatinate, compound with 2-aminoethanol (1:2) (HHPA-2AE)

Time of exposure  (h)	Effect	Control	38.5 mg HHPA-2AE/L	84.7 mg HHPA-2AE/L	186.4 mg HHPA-2AE/L	410.0 mg HHPA-2AE/L	902.0 mg HHPA-2AE/L
			2.3 mg 2AE/L	5.3 mg 2AE/L	11.5 mg 2AE/L	25.3 mg 2AE/L	54.8 mg 2AE/L
			0.42 mg Pt/L	0.94 mg Pt/L	2.17 mg Pt/L	4.86 mg Pt/L	10.9 mg Pt/L
24	N. of mobile daphnids	20	20	20	20	20	20
24	% Immobilisation	0	0	0	0	0	0
48	N. of mobile daphnids	19	20	19	20	20	20
48	% Immobilisation	5	0	5	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The 48-h EC50 value was determined to be > 54.8 mg 2AE L-1 or > 10.9 mg Pt L-1.

Executive summary:

A 48-h acute immobilisation Daphnia magna study is available for dihydrogen hexahydroxyplatinate, compound with 2-aminoethanol (1:2). The study is reliable without restrictions, being GLP-compliant and having followed standard test guidelines (OECD 202 and EU Method C.2).

A semi-static test system was followed, where test solutions were renewed after 24 hours. Five test concentrations were used, and these were analysed separately for platinum and 2-aminoethanol (2AE) using acid digestion with ICP-OES and LC-MS/MS, respectively, at the start and at each renewal until the end of the test. The mean measured concentration of the freshly prepared highest test concentration was found to be 11.0 mg Pt/L, and 10.7 mg Pt/L for the aged test solution. The mean measured concentration of 2AE in the freshly prepared highest test concentration was found to be 53.5 mg/L for 2AE, which corresponded to 55.2 % of nominal. The mean measured concentration of the aged highest test solution was 56.0 mg/L for 2AE, corresponding to 57.8 % of nominal. The measured concentration of the test item could not be based on either of the two measured components, as the test item in its original form was not present anymore in the test solutions. Hence, results were based on mean measured 2AE and Pt concentrations, and nominal concentrations of the test item. All validity criteria were met.

The 48-hour NOEC was estimated to be ≥ 902 mg test item/L (nominal) (corresponding to ≥ 54.8 mg 2AE/L and ≥ 10.9 mg Pt/L), and the 48-h EC50 and EC10 values were both estimated to be > 902 mg test item/L (nominal) (corresponding to > 54.8 mg 2AE/L and > 10.9 mg Pt/L).

Description of key information

The 48-h EC50 value was determined to be > 54.8 mg 2-aminoethanol L^-1or > 10.9 mg Pt L^-1.

Additional information

A 48-h acute immobilisationDaphnia magnastudy is available for dihydrogen hexahydroxyplatinate, compound with 2-aminoethanol (1:2) following OECD guideline 202 and EU Method C.2 (Simon 2016). A semi-static test system was followed, where test solutions were renewed after 24 hours. Five test concentrations were used, and these were analysed separately for platinum and 2-aminoethanol using acid digestion with ICP-OES and LC-MS/MS, respectively, at the start and at each renewal until the end of the test. The mean measured concentration of the freshly prepared highest test concentration was found to be 11.0 mg Pt L^-1, and 10.7 mg Pt L^-1for the aged test solution. The mean measured concentration of 2-aminoethanol in the freshly prepared highest test concentration was found to be 53.5 mg L^-1for 2-aminoethanol, which corresponded to 55.2 % of nominal. The mean measured concentration of the aged highest test solution was 56.0 mg L^-1for 2-aminoethanol, corresponding to 57.8 % of nominal. The measured concentration of the test item could not be based on either of the two measured components, as the test item in its original form was not present anymore in the test solutions. Hence, results were based on mean measured 2-aminoethanol and Pt concentrations, and nominal concentrations of the test item. All validity criteria were met. The 48-hour NOEC was estimated to be ≥902 mg test item L^-1(corresponding to ≥54.8 mg 2-aminoethanol L^-1and ≥10.9 mg Pt L^-1), and the 48-h EC50 and EC10 values were both determined to be > 902 mg test item L^-1 (corresponding to > 54.8 mg 2-aminoethanol L^-1and > 10.9 mg Pt L^-1).