Di(tetraethylammonium)hexahydroxoplatinate(IV)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

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 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

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
- Content: 10.94 % Pt
- Form: liquid
- Appearance: orange liquid

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 15.4, 27.8, 50.0, 90.0 and 162 mg Pt-TEAH/L; 1.69, 3.04. 5.47, 9.85 and 17.7 mg Pt/L (nominal)
- Sampling method: Not stated. One set of samples was filtered through a 0.45 µm PES filter before measurement. The other set of samples was measured without filtration
- Sample storage conditions before analysis: For analysis of Pt, the samples were acidified with HNO3 and immediately stored at 4°C until analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solutions were prepared before test start and media renewal by dissolving test item in dilution water and stirring for two hours. The stock solutions were diluted with test water freshly before use to obtain the test concentrations.
- Controls: Dilution water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): None
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): None
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): None.
- Extraction recovery (indicate if results are corrected or not for recoveries): Pt: 87.1-113%
- Method of confirmation of identity of measured compound:
ICP-OES Pt detection at wavelengths of 203.646 nm, 214.424 nm.
ICP-MS PT detection performed at the isotopes 194Pt, 195Pt and 196Pt.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Strain: Teleostei, Cyprinidae
- Source: Test facility bred
- Age at study initiation (mean and range, SD): Not stated
- Length at study initiation (length definition, mean, range and SD): 1.5 - 1.7 cm
- Weight at study initiation (mean and range, SD): 0.039 - 0.054 g
- Method of breeding: Not stated
- Maintenance of the brood fish: Not stated

ACCLIMATION
- Acclimation period: at least 9 days
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food during acclimation: Live brine shrimp (Artemia salina) nauplii and ground flake food TetraMin.
- Feeding frequency during acclimation: shrimp - ad libitum, flake food – once daily
- Health during acclimation (any mortality observed): Mortality of the batch was less than 5 % in the week preceding the start of the study.

FEEDING DURING TEST
- Food type: None
- Amount: None
- Frequency: None

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

23.0 - 23.2 °C

pH:

7.69 - 8.17

Dissolved oxygen:

98 - 106 %

Salinity:

not applicable

Nominal and measured concentrations:

Nominal concentration: 1.69, 3.04, 5.47, 9.85 and 17.7 mg Pt/L, equivalent to 15.4, 27.8, 50.0, 90.0 and 162 mg test item/L
Measured concentrations: The geometric mean of the measured test item concentration over the test period were between 96.2 and 108.9 % of nominal concentrations. In accordance with OECD 203 the effect assessment was based on the nominal concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel: 3-L glass vessels filled with 2 L test medium
- Aeration: slightly aerated via glass capillaries
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Purified drinking water
- Water quality: Dissolved oxygen concentration, pH-value and temperature were measured directly before adding the fish and afterwards once per day in each test vessel. TOC was measured at the beginning of the exposure in the dilution water.

OTHER TEST CONDITIONS
- Photoperiod: 12 h light and 12 h dark
- Light intensity: 10 – 20 µE/m²/s, equivalent to 540-1000 lux

EFFECT PARAMETERS MEASURED: Fish were observed every 24 hours for symptoms of intoxication and mortality.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: about 1.8
- Range finding study: A non-GLP Range Finder was performed with the test item in order to determine the concentration range which should be applied in the definitive test. In the range finder test three nominal concentrations of 5.0, 50.0 and 500 mg test item/L, equivalent to 0.547, 5.47 and 54.7 mg Pt/L, were applied. The test was performed with five individual specimens per treatment and control.
Within the first 3 hours, all fish in the highest test concentration of 500 mg test item/L (54.7 mg Pt/L) died. After 24 hours, one fish was dead in the second test concentration of 50.0 mg test item/L (5.47 mg Pt/L). No fish died in the control and the lowest test concentration of 5.0 mg test item/L (0.547 mg Pt/L).
- Test concentrations: 15.4, 27.8, 50.0, 90.0 and 162 mg test item/L, equivalent to 1.69, 3.04. 5.47, 9.85 and 17.7 mg Pt/L.
- Results used to determine the conditions for the definitive study: range finder study

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

13.2 mg/L

Nominal / measured:

nominal

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

120.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

12.4 mg/L

Nominal / measured:

nominal

Conc. based on:

element

Remarks:

Pt

Basis for effect:

mortality (fish)

Details on results:

Within the first 24 hours after test start, all fish in the highest concentration of 17.7 mg Pt/L (162 mg test item/L) died. No fish died in the four lower test concentrations of 1.69, 3.04. 5.47 and 9.85 mg Pt/L (15.4, 27.8, 50.0 and 90.0 mg test item/L), respectively. In addition, no fish died in the control.
In the test concentration of 9.85 mg Pt/L (90.0 mg test item/L) two fish showed sublethal effects at 54 hours after application, which could however not be observed anymore at 69 hours after application.

Reported statistics and error estimates:

The LC10, LC50 values and 95 % confidence intervals were calculated using Probit-analysis.
The results were calculated using the nominal concentrations of the test item since recovery rates deviate less than ± 20% of nominal concentrations.
The LC50, the confidence limits (95%) and the slope of the curve were estimated using appropriate statistical methods such as the classical maximum likelihood methods for fitting probit or logit models. All statistics were calculated by using ToxRat® Professional 3.3.0.

Sublethal observations / clinical signs:

Table: Cumulative mortality during the test period of 96 h

		Nominal concentration [mg Test item/L]
		Control		15.4		27.8		50.0		90.0		162
		Nominal concentration [mg Pt/L]
		Control		1.69		3.04		5.47		9.85		17.7
Fish introduced		7		7		7		7		7		7
h of application		Cumulative mortality [n]
0		0.0		0.0		0.0		0.0		0.0		0.0
3		0.0		0.0		0.0		0.0		0.0		0.0
6		0.0		0.0		0.0		0.0		0.0		0.0
21		0.0		0.0		0.0		0.0		0.0		100.0
30		0.0		0.0		0.0		0.0		0.0		100.0
45		0.0		0.0		0.0		0.0		0.0		100.0
54		0.0		0.0		0.0		0.0		0.0		100.0
69		0.0		0.0		0.0		0.0		0.0		100.0
78		0.0		0.0		0.0		0.0		0.0		100.0
93		0.0		0.0		0.0		0.0		0.0		100.0
96		0.0		0.0		0.0		0.0		0.0		100.0

-: no mortality detected

 

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 of di(tetraethylammonium)hexahydroxoplatinate(IV) was determined to be 120.5 mg test item/L, corresponding to 13.2 mg Pt/L.

Executive summary:

A 96-h acute Danio rerio study was conducted for di(tetraethylammonium)hexahydroxoplatinate(IV). The study is reliable without restrictions, being GLP-compliant and having followed standard test guidelines (OECD 203 and EU Method C.1).

A semi-static test procedure was followed. Five test concentrations were used and these were analysed for platinum with ICP-OES and ICP-MS at the start and at each renewal until the end of the test, respectively. The geometric mean of the measured test item concentration over the test period were between 96.2 and 108.9 % of nominal concentrations. In accordance with OECD 203 the effect assessment was based on the nominal concentrations. All validity criteria were met. Di(tetraethylammonium)hexahydroxoplatinate(IV) had a statistically significant effect on the mortality of the fish in the highest test concentrations. The 96-hour LC₅₀was calculated to be 13.2 mg Pt/L, corresponding to 120.5 mg test item/L.

Description of key information

The 96-hour LC₅₀ of di(tetraethylammonium)hexahydroxoplatinate(IV) was determined to be 120.5 mg test item/L, corresponding to 13.2 mg Pt/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

120.5 mg/L

Additional information

A 96-h acute Danio rerio study was conducted for di(tetraethylammonium)hexahydroxoplatinate(IV). The study is reliable without restrictions, being GLP-compliant and having followed standard test guidelines (OECD 203 and EU Method C.1). A semi-static test procedure was followed. Five test concentrations were used and these were analysed for platinum with ICP-OES and ICP-MS at the start and at each renewal until the end of the test, respectively. The geometric mean of the measured test item concentration over the test period were between 96.2 and 108.9 % of nominal concentrations. In accordance with OECD 203 the effect assessment was based on the nominal concentrations. All validity criteria were met. Di(tetraethylammonium)hexahydroxoplatinate(IV) had a statistically significant effect on the mortality of the fish in the highest test concentrations. The 96-hour LC₅₀was calculated to be 13.2 mg Pt/L, corresponding to 120.5 mg test item/L.