complex mixture from biological origin

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

53 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 212 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 212 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

not determinable

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

ca. 25 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: nominal concentration analytically verified

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: nominal concentration analytically verified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 85 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: nominal concentration analytically verified

Duration:

96 h

Dose descriptor:

LC50

Remarks:

24 to 96 hours

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 106 - 212 mg/L

Duration:

96 h

Dose descriptor:

LC50

Remarks:

24 to 96 hours

Effect conc.:

85 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 50.0 - 100

Details on results:

- Behavioural abnormalities: please see table 1 in section “any other details on results incl. tables”.
- Observations on body length and weight: not described
- Mortality of control: none
- Other adverse effects control: not reported
- Abnormal responses: please see table 1 in section “any other details on results incl. tables”.
- Effect concentrations exceeding solubility of substance in test medium: not applicable

Results with reference substance (positive control):

not applicable (no reference item is recommended for this test according to the guidelines)

Validity criteria fulfilled:

yes

Conclusions:

In this study, the test item was found to cause lethal effects to zebrafish after 96 hours at a nominal test item concentration of 106 mg/L and higher. The LC50 after 96 hours was 180 (106 – 212) mg/test item/L, corresponding to 85.0 (50.0 – 100) mg solid content/L. The LC0 and LC100 after 96 hours was 53.0 and >212 mg test item/L, corresponding to 25.0 and >100.0 mg solid content/L (nominal concentrations).

Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2017-02-27 to 2017-03-17, with a definitive exposure phase from 2017-03-13 to 2017-03-17 at the test facility.

An acute toxicity test under semi-static conditions with daily renewal of the test media was conducted with the nominal test item concentrations of 13.3 - 26.5 - 53.0 - 106 - 212 mg/L (factor 2), corresponding to a nominal solid content of 6.28 – 12.5 – 25.0 – 50.0 – 100 mg/L. Duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. Water quality parameters (temperature, pH-value and oxygen-saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits. The determination of the test concentrations was carried out via LC‑MS/MS. Evaluation was done from freshly prepared media after 0 and 72 hours and from the corresponding 24 hours aged test media after 24 and 96 hours. The measured concentrations in freshly prepared media were in the range of 85 and 102 % of the nominal values. In corresponding 24 hours aged test media, the measured concentrations were between 83 and 113 % of the nominal values. Therefore, all effect values are based on nominal

concentrations of the test item. All validity criteria of the test guideline were met. The results of the study are summarized in the table below.

LC-Values (3 – 96 hours) of the Test Item

Based on nominal concentrations of the test item and the solid content [mg/L]

Test duration [h]	Test item [mg/L]		Solid content [mg/L]
	LC50	Confidence interval	LC50	Confidence interval
3	> 212	n.a.	>100	n.a.
24	180	(106 – 212)	85.0	(50.0 – 100)
48	180	(106 – 212)	85.0	(50.0 – 100)
72	180	(106 – 212)	85.0	(50.0 – 100)
96	180	(106 – 212)	85.0	(50.0 – 100)
LC100= Lowest test item concentration with 100 % mortality after 96 h	> 212		> 100
LC0= Highest test item concentration with 0 % mortality after 96 h	53.0		25

This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.

Description of key information

The LC50 after 96 hours was 180 (106 – 212) mg/test item/L, corresponding to 85.0 (50.0 – 100) mg solid content/L.

This information is used in a read-across approach in the assessment of the target substance. For justification of read-across please refer to the attached read-across report (see IUCLID section 13).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

85 mg/L

Additional information

The acute toxicity of the test item to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2017-02-27 to 2017-03-17, with a definitive exposure phase from 2017-03-13 to 2017-03-17 at the test facility.

An acute toxicity test under semi-static conditions with daily renewal of the test media was conducted with the nominal test item concentrations of 13.3 - 26.5 - 53.0 - 106 - 212 mg/L (factor 2), corresponding to a nominal solid content of 6.28 – 12.5 – 25.0 – 50.0 – 100 mg/L. Duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. Water quality parameters (temperature, pH-value and oxygen-saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits. The determination of the test concentrations was carried out via LC‑MS/MS. Evaluation was done from freshly prepared media after 0 and 72 hours and from the corresponding 24 hours aged test media after 24 and 96 hours. The measured concentrations in freshly prepared media were in the range of 85 and 102 % of the nominal values. In corresponding 24 hours aged test media, the measured concentrations were between 83 and 113 % of the nominal values. Therefore, all effect values are based on nominal

concentrations of the test item. All validity criteria of the test guideline were met. The results of the study are summarized in the table below.

LC-Values (3 – 96 hours) of the Test Item

Based on nominal concentrations of the test item and the solid content [mg/L]

Test duration [h]	Test item [mg/L]		Solid content [mg/L]
	LC50	Confidence interval	LC50	Confidence interval
3	> 212	n.a.	>100	n.a.
24	180	(106 – 212)	85.0	(50.0 – 100)
48	180	(106 – 212)	85.0	(50.0 – 100)
72	180	(106 – 212)	85.0	(50.0 – 100)
96	180	(106 – 212)	85.0	(50.0 – 100)
LC100= Lowest test item concentration with 100 % mortality after 96 h	> 212		> 100
LC0= Highest test item concentration with 0 % mortality after 96 h	53.0		25

This information is used in a read-across approach in the assessment of the target substance. For justification of read-across please refer to the attached read-across report (see IUCLID section 13).