Reaction mass of 3,4,5-trichlorophthalic acid, strontium salt and 3,4,6-trichlorophthalic acid, strontium salt and 3,4-dichlorophthalic acid, strontium salt and 3,5-dichlorophthalic acid, strontium salt and 3,6-dichlorophthalic acid, strontium salt and 3-chlorophthalic acid, strontium salt and 4,5-dichlorophthalic acid, strontium salt and 4-chlorophthalic acid, strontium salt

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

Study period:

2019-09-11 to 2019-11-13

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)

Version / remarks:

Adopted April 13, 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Limit Test, 100 mg test item/L nominal. The concentrations of the test item strontium dichlorophthalate were analysed in the duplicate test media samples from all test concentrations, in the duplicate control samples and in the additional control and solvent samples, from both sampling times (0 and 48 hours).
- Sampling method: The samples were taken from the biological phase of the study. Duplicate samples from the freshly prepared test medium of the only test concentration and the control were taken at the start of the test. For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentration during the test period, duplicate samples from the test medium of the only test concentration and the control were collected at the end of the test (after 48 hours). All samples were diluted by a factor of 2 with acetonitrile. Additional samples of the control and the dilution solvent were taken at test start and test end without any sample treatment
- Sample storage conditions before analysis: All samples were stored in a freezer (≤ - 20 °C), protected from light, until analysis was performed. Afterwards the samples were again stored deep frozen (≤ -20 °C) and will be kept stored up to the date of the final report.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item was used to prepare a stock solution. 50.01 mg of the test item were dissolved in 50 mL pure water (20 minutes ultrasonication) to obtain a stock solution of approximately 1 g test item/L. Appropriate amounts of the stock solution were diluted with acetonitrile / test water (1/1, v/v) to obtain standard solutions in the range from 10 – 100 mg test item/L. Exact data were documented in the raw data.
- Eluate: N/A
- Differential loading: N/A
- Controls: In the control, test water was used without addition of the test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N/A
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): N/A
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Straus)
- Strain/clone: clone 5
- Justification for species other than prescribed by test guideline: N/A
- Age at study initiation (mean and range, SD): From 1.5 to 20.5 hours old
- Method of breeding: The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium. The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.
- Source: The daphnids introduced in the test were taken from ibacon's in house laboratory culture
- Feeding during test : No
- Food type: N/A
- Amount: N/A
- Frequency: N/A
Reference Item: For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.

ACCLIMATION
Acclimation was not necessary, since the test was performed in the same medium as the culturing

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

2.5 mmol/L (= 250 mg/L) as CaCO3

Test temperature:

19.9 to 20.6 °C at test start;
20.0 °C at test end

pH:

7.8 to 7.9 at test start;
7.9 to 8.0 at test end

Dissolved oxygen:

8.9 to 9.0 mg/L at test start;
9.0 mg/L at test end

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: At the start of the test 104 % of the nominal test concentrations were found (average of the only test concentration). After 48 hours test duration, 104 % of the nominal value was determined (average of the only test concentration). During the test the daphnids were exposed to a mean of 104 % of nominal.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers of 100 mL volume containing approximately 60 mL of test medium. The test vessels were covered with a lid to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions.
- Type (delete if not applicable): open / closed : Open
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): Static, no flow-through
- Renewal rate of test solution (frequency/flow rate): N/A
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): N/A
- Biomass loading rate: N/A

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised water
- Test medium: Elendt "M4"
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light - 8 h dark
- Light intensity: 240 to 480 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility. The mobility of the daphnids was determined by visual observation after 24 and 48 hours. Those animals that are not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile (even if they could still move their antennae). The mobility of the daphnids was determined in a semi-static 48-hour test by visual observation after 24 and 48 hours.

VEHICLE CONTROL PERFORMED: No

RANGE-FINDING STUDY
- Test concentrations: 100 mg test item/L
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: No
- Observations on body length and weight: No
- Mortality of control: No
- Other adverse effects control: No
- Immobilisation of control: Two immobile animals were observed in the control
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

In the most recent test with the reference item potassium dichromate performed in February 2019 (study code 139881220), the EC50 after 24 hours was determined to be 0.932 mg test item/L, indicating that the sensitivity of the daphnids was consistent with the level proposed by the OECD 202 guideline (EC50-24 h between 0.6 and 2.1 mg potassium dichromate/L).

Reported statistics and error estimates:

No statistical analysis was performed.
The EC50 could not be quantified due to the absence of toxicity of the test item and were determined to be higher than the highest test concentration. The NOEC and the LOEC were determined directly from the raw data.

Validity Criteria of the Study:

Control Immobilisation Rate: 10 %; Furthermore, no daphnid showed signs of disease or stress; thus, the validity criterion was met.

 

Dissolved Oxygen Concentration: Was 9.0 mg O2/L in in all treatment groups at the end of the test; thus, the validity criterion was met.

 

 

Biological Results:

 

Signs of Intoxication after 48 Hours:

After 48 hours of exposure no immobile animal was observed at the concentration of 100 mg test item/L, the only concentration tested. Two immobile animals were observed in the control (see table 1 and 2).

 

 

Table 1: Influence of Strontium Dichlorophthalate on the Mobility ofDaphnia magnaafter 24 and 48 hours of exposure. The values are presented in nominal concentrations as measured concentrations were 104 % of the nominal concentrations. 

Endpoint [mg test item/L]	24 hours	48 hours
EC50	> 100	> 100
95 %CI	n.d.	n.d.
EC20	> 100	> 100
95 %CI	n.d.	n.d.
EC10	> 100	> 100
95 %CI	n.d.	n.d.
NOEC	≥ 100	≥ 100
LOEC	> 100	> 100

n.d.: not determinable NOEC and LOEC were determined directly from the raw data.

 

 

Table 2: Observations on the Mobility of Daphnia magna during exposure to Strontium Dichlorphthalate.

Nominal concentration [mg test item/L]	No. of Daphnia tested	No. of immobilised Daphnia after		of immobilised Daphnia after		Observations
		24 h	48 h	24 h	48 h	24 h	48 h
Control	20	0	2	0	10	0	0
100	20	0	0	0	0	0	0

 

Analytical Results 

Table 3: Summary of Analytical Results

Sample description	Sample age		Concentration found^1		Concentration calculated^1	Concentration nominal^1	% of nominal
mg test item/L	Day	Hour	mg test item/L	Dilution factor	mg test item/L	mg test item/L
Control	0	0	<LOD	2	n.a.	0.000	n.a.
Control	0	0	<LOD	2	n.a.	0.000	n.a.
Control	2	48	<LOD	2	n.a.	0.000	n.a.
Control	2	48	<LOD	2	n.a.	0.000	n.a.
100	0	0	51.584	2	103.167	100.000	103
100	0	0	52.509	2	105.018	100.000	105
100	2	48	50.641	2	101.282	100.000	101
100	2	48	53.450	2	106.899	100.000	107
						Mean value (n=4)	104
						RSD (n=4)	2

 

¹The tabulated results represent rounded results calculated on the exact raw data

LOD: Limit of Detection = 3 mg test item/L

RSD: relative standard deviation per treatment group

n: number of analysed samples

n.a.: not applicable

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test conducted as Limit test according to the OECD guideline 202 Daphnia magna were exposed to 100 mg/L strontium dichlorophthalate. No effect was observed.

Executive summary:

The 48-hr-acute toxicity of strontium dichlorophthalate to Daphnia magna was studied under static conditions. Test species were exposed to control and test chemical at nominal concentration of 100 mg test item/L for 48 hr. Mortality/immobilisation and sublethal effects were observed daily. No toxic effect was observed.

 

Based on the results of this study, strontium dichlorophthalate would be classified as not toxic to Daphnia magna in accordance with the classification system of the EU.

 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^st instar): <24 h

Test Type (Flowthrough, Static, Static Renewal): Static

Summary of Biological Results
Nominal Concentration	% of immobilised daphnids after
[mg test item/L]	24 hours	48 hours
Control	0	0
100 mg test item/L	0	0
EC50[mg test item/L]:	> 100	> 100
95 % CI [mg test item/L]:	n.d.	n.d.
EC20[mg test item/L]:	>100	> 100
95 % CI [mg test item/L]:	n.d.	n.d.
EC10[mg test item/L]:	> 100	> 100
95 % CI [mg test item/L]:	n.d.	n.d.
NOEC [mg test item/L]:	≥100	≥100
LOEC [mg test item/L]:	> 100	> 100

 

Endpoint(s) Effected: mobility, mortality

Description of key information

The toxicity of strontium dichlorophthalate was assessed in a 48-hr acute toxicity test (limit test design) with Daphnia magna according to OECD test guideline 202 and under GLP.

No effects were observed in the test. The highest concentration tested was 100 mg/L.