Reaction mass of calcium dihydroxide and calcium peroxide

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

Study period:

19/03/2018 - 23/03/2018

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:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23

Version / remarks:

2000

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Remarks:

not feasible.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A two-day period of magnetic stirring was applied to obtain the maximum dissolution of the test item in medium. Thereafter, the Water Accommodated Fractions (WAFs) were collected by means of filtration through a 0.45 µm membrane filter (Supor®, Pall Corporation). pH of these WAFs was adjusted using 1N HCl (Merck, Darmstadt, Germany) from 8.3-11.3 to 7.9-8.0 for the range-finding test and from 11.3 to 8.1 for the final test.
- Differential loading: Preparation of test solutions started with loading rates individually prepared at 1.0, 10 and 100 mg/L for the range-finding test and at 100 mg/L for the final (limit) test.
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No. All test solutions were clear and colorless at the end of the preparation procedure.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Strain: Cyprinus carpio, Teleostei, Cyprinidae, Linnaeus, 1758
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): Range-finding test: 3.1 ± 0.4 cm; Final test: 2.5 ± 0.2 cm.
- Weight at study initiation (mean and range, SD): Range-finding test: 0.42 ± 0.13 g; Final test: 0.16 ± 0.04 g

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions: same as test
- Type and amount of food during acclimation: pelleted fish food (Essence (300-500 μm), Coppens International bv, Helmond, The Netherlands)
- Feeding frequency during acclimation: daily
- Health during acclimation: In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

FEEDING DURING TEST
- No feeding from 24 hours prior to the test and during the total test period.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

180 mg/L CaCO3

Test temperature:

21.2 ± 0.5

pH:

7.7 ± 0.3

Dissolved oxygen:

7.8 ± 0.9

Nominal and measured concentrations:

100 mg/L WAF.

Details on test conditions:

TEST SYSTEM
- Test vessel: 5L glass vessels
- Material, size, headspace, fill volume: glass, 5L, containing 4L of test solution
- Aeration: no
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.28 g fish/Litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
- Culture medium different from test medium: no
- Intervals of water quality measurement: water quality parameters were kept within the optimum limits for the respective fish species.

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality and other effects at 3, 24, 48, 72 and 96 hours following the start of exposure. In addition, every afternoon from day 0 to observe for any dead or severely distressed fish. Dissolved oxygen content, pH and temperature were measured daily in all vessels from the start of the test (day 0).

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1, 10, 100 mg/L loading rates.
- Results used to determine the conditions for the definitive study: No mortality or clinical effects were observed at any of the test concentrations during the test period. Therefore, the expected LC50 was above the concentration obtained in a WAF prepared at 100 mg/L. Therefore, a limit test was performed.

Reference substance (positive control):

yes

Remarks:

pentachlorophenol (PCP, SIGMA, Art. P2604, Batch MKBW8316V)

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: No mortality or clinical effects were observed in the WAF and the control during the test period.
- Mortality of control: no.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: PCP induced no lethal effects in carp at 0.10 mg/L. 100% mortality observed at 0.22 and 0.46 mg/L. This effect was already reached within 24 hours of exposure.
- LC50: The 96h-LC50 for carp exposed to PCP was 0.15 mg/L (95 % confidence interval between 0.10 and 0.22 mg/L).
- Other: The range of the 96h-LC50 for carp is generally between 0.10 and 0.46 mg/L based on historical data of reference tests performed approximately every 3 months from April 1988 until the end of 2000, and annually since then.

Sublethal observations / clinical signs:

Table 1. Incidence of Mortality and Total Mortality during the Range-Finding Test.

Test item Loading rate^1 (mg/L)	Initial number of fish	Cumulative mortality					Total Mortality (%)
		2¼h	24h	48h	72h	96h
1.0	3	0	0	0	0	0	0
10	3	0	0	0	0	0	0
100	3	0	0	0	0	0	0

¹: WAF prepared at the given loading rate.

 

Table 2. Incidence of Mortality and Total Mortality during the Limit Test.

Test item Loading rate^1 (mg/L)	Initial Number of fish	Cumulative mortality					Total Mortality (%)
		3h	24h	48h	72h	96h
Control	7	0	0	0	0	0	0
100	7	0	0	0	0	0	0

¹: WAF prepared at the given loading rate.

 

Table 3. Effect Parameters

Parameter	Loading rate test item (mg/L)
24h-LL50	>100
48h-LL50	>100
72h-LL50	>100
96h-LL50	>100

 

Table 4. pH-Values and Dissolved Oxygen Concentrations (mg/L) during the Limit Test

Test item Loading rate^1 (mg/L)	Day 0		Day 1		Day 2		Day 3		Day 4
	pH^2	O2	pH	O2	pH	O2	pH	O2^3	pH	O2
Control	8.0	8.8	7.9	7.7	7.9	7.4	8.1	6.2	7.6	6.5
100	8.1	9.1	7.8	7.9	7.8	7.9	7.8	6.9	7.4	7.1

¹ WAF prepared at the given loading rate; ² pH of culture medium was: 7.9; ³ O2 was measured again in the afternoon and was 5.9 mg/L for the control and 6.7 mg/L for the WAF.

Table 5. Temperatures (°C) Measured during the Limit Test

Test item Loading rate^1 (mg/L)	Day 0^2	Day 1	Day 2	Day 3	Day 4
Control	23	21	21	21	21
100	22	21	21	21	21

¹ WAF prepared at the given loading rate; ² Temperature of culture medium was: 23°C.

Validity criteria fulfilled:

yes

Remarks:

No mortality was observed in the control at the end of the test; test conditions were maintained constant throughout the test; the dissolved oxygen concentration has been at least 60% of the air saturation value throughout the test (>5 mg/L at 22°C).

Conclusions:

The 96h-LL50 of the test item in carp was found to be higher than 100 mg/L.

Executive summary:

An acute toxicity test on Carp (Cyprinus carpio) was performed in accordance with OECD Guideline 203 / EU method C.1, under GLP conditions. Based on the results obtained in a preliminary range-finding test where no toxic response was observed, a Limit Test was carried out using only one test concentration (100 mg/L) and one control group in a static system. A Water Accommodated Fraction (WAF) was prepared at a loading rate of 100 mg/L, and the pH was adjusted from 11.3 to 8.1. Seven fish per group were exposed to the test item WAF or the control for 96 h. All validity criteria were met. No mortality or clinical effects were observed in the WAF and the control during the test period. The 96h-LL50 for carp exposed to the test item was found to be >100 mg/L based on loading rate.

Description of key information

Key study. Method according to OECD 203, GLP study. The test item has a 96h-LL50 > 100 mg/L in fish.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

An acute toxicity test on Carp (Cyprinus carpio) was performed in accordance with OECD Guideline 203 / EU method C.1, under GLP conditions. Based on the results obtained in a preliminary range-finding test where no toxic response was observed, a Limit Test was carried out using only one test concentration (100 mg/L) and one control group in a static system.A Water Accommodated Fraction (WAF) was prepared at a loading rate of 100 mg/L, and the pH was adjusted from 11.3 to 8.1.Seven fish per group were exposed to the test item WAF or the control for 96 h. All validity criteria were met.No mortality or clinical effects were observed in the WAF and the control during the test period. The 96h-LL50 for carp exposed to the test item was found to be >100 mg/L based on loading rate.