Di(tetraethylammonium)hexahydroxoplatinate(IV)

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)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: 3341/00-F9
- Expiration date of the lot/batch: August 06, 2020
- Purity test date: Certificate of analysis dated August 06, 2019
- Content: 10.94 % Pt
- Form: liquid
- Appearance: clear orange liquid

Analytical monitoring:

yes

Details on sampling:

Samples with five different nominal concentrations of 4.27, 9.39, 20.7, 45.5 and 100 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).

Vehicle:

no

Details on test solutions:

For preparation of test solutions, at first a stock solution was prepared by adding an appropriate amount of the test item to 1 L dilution water. This stock solution served as highest test concentration (nominal concentration 100 mg/L). The stock solution was stirred for 2 hours at room temperature. Afterwards, the test solution was serially diluted with dilution water to obtain the lower test concentrations.
The test was performed under semi-static test conditions. Test solutions were renewed after 24 hours. The test solutions were freshly prepared before test start and before media renewal.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- 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, at least 3 weeks old, were separated from the stock population by sieving. Batches of 30 to 50 animals were held at room temperature in ca. 1.8 L dilution water for one week. During this week the daphnids were fed daily with an algal suspension (Desmodesmus subspicatus) and ArtemioFluid (JBL). Algae growing in the log-phase were centrifuged and the pellet was re-suspended in a few mL of medium. 5 mL of this suspension was given to 1.8 L Daphnia medium. The water was changed three times 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 specimens.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

1.4 mmol/l

Test temperature:

18.8 - 19.1 °C

pH:

7.94 – 8.22

Dissolved oxygen:

8.09 mg/L - 9.43 mg/L (91.1 – 105 % of air saturation)

Conductivity:

247 µS/cm

Nominal and measured concentrations:

nominal concentrations: 4.27, 9.39, 20.7, 45.5 and 100 mg test item/L (0.467, 1.03, 2.26, 4.98, 10.9 mg Pt/L)
mean measured concentrations: 0.475, 1.05, 2.27, 5.11 and 11.0 mg Pt/L (between 100 - 103% of nominal concentrations)

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, Cu-reduced drinking water was used as holding- and dilution water. The purification includes filtration with activated charcoal and aeration. To avoid copper contamination, plastic water pipes are used for the testing facilities.
Hardness of the Cu-reduced water is below the recommended range (>140 mg/L as CaCO3) for Daphnia magna indicated in the guideline. Therefore, the Cu-reduced dilution water is enriched with an appropriate amount of CaCl¬2 to increase the hardness, resulting in values of about 250 – 350 mg CaCO3/L.
- Total Hardness: 1.4 mmol/L
- Metals: Cd, Cr, Cu, Mn, Ni, Pb and Zn < LOQ; Fe 1.91 - 2.04 μg/L
- Chlorine: 0.03 mg/L
- Alkalinity: 1.4 mmol/L
- Conductivity: 247 μS/cm
- Intervals of water quality measurement: oxygen concentration, pH and temperature of the test solutions were checked at test start, at media renewal and at test end.

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours dark
- Light intensity: 740 - 889 lux (corresponding to 9.9 – 11.9 μE/(m² *s))

EFFECT PARAMETERS MEASURED
Immobile daphnids were determined after 24 h and at the end of the test (48 h). The animals were considered to be immobile when they were not able to swim within 15 seconds after gentle agitation of the test vessels. Any abnormalities in appearance and behaviour were also recorded.

RANGE-FINDING STUDY
- Test concentrations: 1.0, 10.0 and 100 mg/L test item
- Results used to determine the conditions for the definitive study:
The non-GLP range-finding test was conducted with four replicates with five daphnids per beaker under semi-static test conditions with renewal of test solution once after 24 hours.
At test end, no mortality occurred in the control. In the lowest test concentration of 1.0 mg/L 15% of the introduced daphnid died. At the second and third test concentration of 10.0 and 100 mg/L, 25% of the introduced daphnids died. In addition, daphnids in the highest test concentration were less active than daphnids in the control. They were lying on the ground or hanging on the surface of the water and they were barely moving, only when gently touched with a pipette. However, the observed effects were statistically not significant.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

174.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

19.1 mg/L

Nominal / measured:

nominal

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

9.39 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.03 mg/L

Nominal / measured:

nominal

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

20.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

2.27 mg/L

Nominal / measured:

nominal

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mobility

Details on results:

No significant signs of disease or stress like discolouration or abnormal behaviour were observed in any replicate.

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:      Mobile and immobile daphnids after 48 h and percent immobilization

               compared to the controls. Concentrations given as nominal test item and Pt concentrations.

Nominal concentration Test item [mg/L]	Nominal concentration Pt [mg/L]	Total Introduced	Mobile	Immobile	% Immobility
Control	Control	20	20	0	0.0
4.27	0.467	20	20	0	0.0
9.39	1.03	20	20	0	0.0
20.7	2.26	20	16	4	20.0
45.5	4.98	20	15	5	25.0
100	10.9	20	14	6	30.0

Validity criteria fulfilled:

yes

Conclusions:

The EC50 was calculated to be 174.5 mg test item/L (19.1 mg Pt/L), the NOEC was set to 9.39 mg test item/L (1.03 mg Pt/L).

Executive summary:

A 48-h acute immobilisation Daphnia magna study is available for Di(tetraethylammonium)hexahydroxoplatinate(IV) (aqueous solution). 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 for platinum with ICP-OES atat beginning of the test, before and after media renewal at 24 hours and at test end.

Concentrations of freshly prepared test media showed recovery rates between 96.6 and 108 % of nominal concentrations. Concentrations in aged test media showed recovery rates between 96.4 and 111 % of nominal concentrations. During incubation in the test, test concentrations remained stable with 90.5 to 115 % of initial concentrations. The measured concentrations were within a range of 80 – 120 % of nominal concentrations and remained stable during the test. Thus, the evaluation of effects was based on the nominal concentrations.

All validity criteria were met.

The test item had a statistically significant effect on the immobilization of the daphnids in the three highest test concentrations of 20.7, 45.5 and 100 mg test item /L. The EC₅₀was calculated to be 174.5 mg test item/L (19.1 mg Pt/L), the NOEC was set to 9.39 mg test item/L (1.03 mg Pt/L).

Description of key information

The test item had a statistically significant effect on the immobilization of the daphnids in the three highest test concentrations of 20.7, 45.5 and 100 mg test item /L. The EC₅₀was calculated to be 174.5 mg test item/L (19.1 mg Pt/L), the NOEC was set to 9.39 mg test item/L (1.03 mg Pt/L).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

174.5 mg/L

Additional information

A 48-h acute immobilisation Daphnia magna study is available for Di(tetraethylammonium)hexahydroxoplatinate(IV) (aqueous solution). 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 for platinum with ICP-OES at beginning of the test, before and after media renewal at 24 hours and at test end.

Concentrations of freshly prepared test media showed recovery rates between 96.6 and 108 % of nominal concentrations. Concentrations in aged test media showed recovery rates between 96.4 and 111 % of nominal concentrations. During incubation in the test, test concentrations remained stable with 90.5 to 115 % of initial concentrations. The measured concentrations were within a range of 80 – 120 % of nominal concentrations and remained stable during the test. Thus, the evaluation of effects was based on the nominal concentrations.

All validity criteria were met.

The test item had a statistically significant effect on the immobilization of the daphnids in the three highest test concentrations of 20.7, 45.5 and 100 mg test item /L. The EC₅₀was calculated to be 174.5 mg test item/L (19.1 mg Pt/L), the NOEC was set to 9.39 mg test item/L (1.03 mg Pt/L).