Benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-10-01 to 2019-10-15

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:

2004

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

For the analytical measurement a sample from the freshly prepared test medium from the concentration of the closed system was taken at the start of the study. For the determination of the maintenance of the test material concentration during the exposure period a sample was taken from the medium at the end of the exposure period.

Vehicle:

yes

Remarks:

Reconstituted water

Details on test solutions:

A preparation with a test item concentration of 100 mg/L was freshly prepared. For that purpose, the test item was weighed in a calibrated flask and vehicle, reconstituted water was added. The preparation was stirred with a magnetic stirrer for 1 hour. Then the formulation was passed through a filter membrane (pore size: 0.2 µm). The filtrate was used for the preparation of the different concentrations. The pH was not adjusted.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus
- Age at study initiation: not older 24 h
- Source: IBACON GmbH (Roßdorf, Germany)
- Feeding during test: No

ACCLIMATION
- Acclimation period: No
- Breeding conditions: same as test
- Type and amount of food: unicellular green algae
- Feeding frequency: fed ad libitum with unicellular green algae three times per week
- Health during acclimation: no mortality observed

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

240 - 246 mg CaCO3/L

pH:

7.06 - 7.10

Dissolved oxygen:

8.30 - 8.43 mg/L

Conductivity:

677 – 684 µS/cm

Nominal and measured concentrations:

Nominal concentrations: 0, 0.10, 0.22, 0.48, 1.06, 2.34, 5.15, 11.34 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessel
- Type: open
- Material, size, fill volume: glass, 20 mL, 20 mL
- Volume of solution: 20 mL
- Aeration: No
- Renewal rate of test solution: every 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
- Preparation of dilution water: The water was composited according to ELENDT M4 (1990)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hour light and 8 hour dark regime

EFFECT PARAMETERS MEASURED:
The mobility was determined after 24 and 48 hours

RANGE-FINDING STUDY
- Test concentrations: 0, 0.1, 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes, at 100 mg/L Daphnia showed immobilization

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.25 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

0.52 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no
- Observations on body length and weight: no
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Immobilisation of control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

No effect on the mobility of the daphnia was observed at the two lowest nominal concentrations of 0.10 and 0.22 mg/L. A concentration dependent immobilization was observed at nominal concentrations of 0.48 to 5.15 mg/L.

Table 1: The number of immobilized daphnids per test item concentration

Nominal Test Item Concentration [mg/L]	Number of daphnids immobilized / exposed		% of daphnids immobilized
	24 hours	48 hours	24 hours	48 hours
0.0	0/20	0/20	0%	0%
0.10	0/20	0/20	0%	0%
0.22	0/20	0/20	0%	0%
0.48	0/20	5/20	0%	25%
1.06	8/20	16/20	40%	80%
2.34	15/20	20/20	75%	100%
5.15	20/20	20/20	100%	100%
11.34	20/20	20/20	100%	100%

Table 2: EC₅₀-Values

Test Duration	EC50	95% Confidence Interval
24h	0.52 mg/L	0.41 - 0.65 mg/L
48h	0.25 mg/L	0.20-0.31 mg/L

Validity criteria fulfilled:

yes

Conclusions:

The test item showed acute toxicity towards daphnia magna according to OECD 202. For the test item, the following EC values were determined: 24-h EC50 was 0.52 mg/L and 48-h EC50 was determined to be 0.25 mg/L.

Executive summary:

The objective of this study was to determine the effect of the test item on the immobilization of Daphnia magna according to OECD 202 and Commission Regulation (EU) 2016/266. For this purpose, juvenile daphnids were exposed to different concentrations of the test item (0.10. 0.22, 0.48, 1.06, 2.34, 5.15 and 11.34 mg/L) and a control in an open semi-static system. 20 daphnids, divided into four replicates, each with five animals were used in the test item group and the control group. The daphnids were observed for immobilization after 24 and 48 hours of exposure. The test item preparation (test item groups) and the vehicle (control group) were renewed after 24 hours. Concentrations of the test item in the test medium were analyzed via HPLC with UV-detection directly after each preparation (fresh medium) and 24h after each preparation (old medium). The recovery rates of the test item in fresh medium at the nominal concentrations of 0.48, 1.06, 2.34, 5.15 and 11.34 mg/L were between 62.3 % to 70.0 % and <LOQ for the nominal concentrations of 0.10 and 0.22 mg/L at 0 hours and 24 hours. In the old medium the recovery rates at the nominal concentrations of 2.34, 5.15 and 11.34 mg/L were between 17.6 % to 23.6 % and <LOQ for the nominal concentrations of 0.10, 0.22, 0.48 and 1.06 mg/L at 24 hours and 48 hours. No test item was measured in the control medium. As the test item concentrations could not be maintained >80% of the measured initial concentrations, the EC values were calculated based on the calculated geometric mean concentrations. For measured concentrations <LOQ the concentrations used for calculation of the geometric mean were extrapolated with the mean recovery of the higher concentrations at the same time point where analytical values were >LOQ. No effect on the mobility of the daphnia was observed at the two lowest nominal concentrations of 0.10 and 0.22 mg/L. A concentration dependent immobilization was observed at nominal concentrations of 0.48 to 5.15 mg/L. Under the conditions of the present study, the test item showed a 48h EC50 of 0.25 mg/L for the immobilization of Daphnia magna.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

(Q)SAR

Adequacy of study:

supporting study

Study period:

2019-09-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

Please refer to attached QMRF and QPRF Documents

Qualifier:

according to guideline

Guideline:

other: Guidance on information requirements and chemical safety assessment, Chapter R.6: QSARs and grouping of chemicals

Deviations:

not applicable

Principles of method if other than guideline:

Calculation of Short-Term toxicity in Daphnia. Software used: ECOSAR 1.11 (EPISUITE) for chemical class "Neutral Organics"

GLP compliance:

no

Specific details on test material used for the study:

Smiles: c12c(cccc1)N(OP(N1CCCC1)(N1CCCC1)N1CCCC1)N=N2

Analytical monitoring:

not required

Test organisms (species):

other: Daphnia

Water media type:

freshwater

Remarks on exposure duration:

acute exposure (duration not specified)

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

7.206 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

not specified

--------------------------------------

ECOSAR v1.11 Class-specific Estimations

--------------------------------------

Benzotriazoles

Predicted

ECOSAR Class                Organism           Duration End Pt  mg/L (ppm)

=========================== ================== ======== ======  ==========

Benzotriazoles            : Daphnid            48-hr    LC50    7.206

 

Validity criteria fulfilled:

yes

Conclusions:

Using ECOSAR v1.11 the short-term toxicity 48-h EC50 value for Daphnia was determined to be 7.206 mg/L. The substance is within the applicability domain of the model.

Executive summary:

Using ECOSAR v1.11 the short-term toxicity 48-h EC50 value for Daphnia was determined to be 7.206 mg/L, as no effects up to the limit dose were observed. The substance is within the applicability domain of the model.

The adequacy of a prediction depends on the following conditions:

a) the (Q)SAR model is scientifically valid: the scientific validity is established according to the OECD principles for (Q)SAR validation;

b) the (Q)SAR model is applicable to the query chemical: a (Q)SAR is applicable if the query chemical falls within the defined applicability domain of the model;

c) the (Q)SAR result is reliable: a valid (Q)SAR that is applied to a chemical falling within its applicability domain provides a reliable result;

d) the (Q)SAR model is relevant for the regulatory purpose.

For assessment and justification of these 4 requirements the QMRF and QPRF files were developed and attached to this study record.

 

Description of the prediction Model

The prediction model was descripted using the harmonised template for summarising and reporting key information on (Q)SAR models. For more details please refer to the attached QSAR Model Reporting Format (QMRF) file. 

 

Assessment of estimation domain

The assessment of the estimation domain was documented in the QSAR Prediction Reporting Format file (QPRF). Please refer to the attached document for the details of the prediction and the assessment of the estimation domain.

Description of key information

The test item showed acute toxicity towards daphnia magna according to OECD 202. For the test item, the following EC values were determined: 24-h EC50 was 0.52 mg/L and 48-h EC50 was determined to be 0.25 mg/L (reference 6.1.3 -2).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.25 mg/L

Additional information

The objective of this study was to determine the effect of the test item on the immobilization of Daphnia magna according to OECD 202 and Commission Regulation (EU) 2016/266. For this purpose, juvenile daphnids were exposed to different concentrations of the test item (0.10. 0.22, 0.48, 1.06, 2.34, 5.15 and 11.34 mg/L) and a control in an open semi-static system. 20 daphnids, divided into four replicates, each with five animals were used in the test item group and the control group. The daphnids were observed for immobilization after 24 and 48 hours of exposure. The test item preparation (test item groups) and the vehicle (control group) were renewed after 24 hours. Concentrations of the test item in the test medium were analyzed via HPLC with UV-detection directly after each preparation (fresh medium) and 24h after each preparation (old medium). The recovery rates of the test item in fresh medium at the nominal concentrations of 0.48, 1.06, 2.34, 5.15 and 11.34 mg/L were between 62.3 % to 70.0 % and <LOQ for the nominal concentrations of 0.10 and 0.22 mg/L at 0 hours and 24 hours. In the old medium the recovery rates at the nominal concentrations of 2.34, 5.15 and 11.34 mg/L were between 17.6 % to 23.6 % and <LOQ for the nominal concentrations of 0.10, 0.22, 0.48 and 1.06 mg/L at 24 hours and 48 hours. No test item was measured in the control medium. As the test item concentrations could not be maintained >80% of the measured initial concentrations, the EC values were calculated based on the calculated geometric mean concentrations. For measured concentrations <LOQ the concentrations used for calculation of the geometric mean were extrapolated with the mean recovery of the higher concentrations at the same time point where analytical values were >LOQ. No effect on the mobility of the daphnia was observed at the two lowest nominal concentrations of 0.10 and 0.22 mg/L. A concentration dependent immobilization was observed at nominal concentrations of 0.48 to 5.15 mg/L. Under the conditions of the present study, the test item showed a 48h EC50 of 0.25 mg/L for the immobilization of Daphnia magna.

For supporting information, QSAR was used for the evlaution of the test item.

Using ECOSAR v1.11 the short-term toxicity 48-h EC50 value for Daphnia was determined to be 7.206 mg/L. The substance is within the applicability domain of the model.

The adequacy of a prediction depends on the following conditions:

a) the (Q)SAR model is scientifically valid: the scientific validity is established according to the OECD principles for (Q)SAR validation;

b) the (Q)SAR model is applicable to the query chemical: a (Q)SAR is applicable if the query chemical falls within the defined applicability domain of the model;

c) the (Q)SAR result is reliable: a valid (Q)SAR that is applied to a chemical falling within its applicability domain provides a reliable result;

d) the (Q)SAR model is relevant for the regulatory purpose.

 

For assessment and justification of these 4 requirements the QMRF and QPRF files were developed and attached to this study record.

 

Description of the prediction Model

The prediction model was descripted using the harmonised template for summarising and reporting key information on (Q)SAR models. For more details please refer to the attached QSAR Model Reporting Format (QMRF) file. 

 

Assessment of estimation domain

The assessment of the estimation domain was documented in the QSAR Prediction Reporting Format file (QPRF). Please refer to the attached document for the details of the prediction and the assessment of the estimation domain.