(2-hydroxy-1,1-dimethylethyl)ammonium toluene-4-sulphonate

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:

April. 21. 2020 to September.01.2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

OECD-Guideline for Testing of Chemicals No. 203 (2019)
OECD Series on Testing and Assessment, No. 126, Short Guidance on the Threshold Approach for Acute Fish Toxicity

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

The limit test concentration and the control was analytically verified via HPLC-DAD in the fresh media at the start of exposure and at one renewal interval (0 and 48 hours) and in the corresponding 48-hours aged media after 48 and 96 hours.

Details on sampling:

- Limit concentration 70 mg/L
- The sample was diluted with ultra-pure water prior to analysis.
- All samples were stored at room temperature until start of analysis, if necessary. Prepared samples were stored at room temperature in an autosampler until analysis.

Vehicle:

no

Details on test solutions:

An appropriate amount of the test item was weighed and transferred into a test vessel filled with an appropriate amount of the dilution water. The test media was mixed with an ultraturrax (1 min, 17000 rpm). This procedure was repeated for every water renewal.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Danio rerio (zebrafish)
Vertebrata, Gnathostomata, Pisces, Osteichthyes, Teleostei, Cypriniformes, Cyprinidae
All fish used in the test were gained at Noack Laboratorien GmbH from a single brood stock (supplier: Umweltbundesamt, Schichauweg 58, D-12307 Berlin)

Juvenile fish

Holding was performed in accordance with the laboratory SOPs at the test facility at 23 ± 2 °C and diffuse light (540 – 1000 Lux, photoperiod of 12 to 16 h light / 12 to 8 h dark). The water was changed at least once per week. The dissolved oxygen concentration was more than 80% of the air saturation value. Zebrafish with at least 9 days of acclimatization and mortality < 5% within these days prior to the start of the exposure were used in the test.
No mortality was observed during this time. No disease treatments were administered throughout holding and testing.

Tap water of local origin was used for holding. The water was filtered on activated charcoal and aerated for at least 24 h to remove chlorine.
Nominal water parameters:
Total hardness: 40 - 250 mg CaCO3/L
pH- value: 6.0 - 8.5
Oxygen saturation: ≥ 80 % of air saturation value
Acidity: 0.2 mmol/L (recent measurement 2020-04-27)
Alkalinity: 0.5 mmol/L (recent measurement 2020-04-27)
Conductivity: 144 µS/cm (recent measurement 2020-04-27)

Food was provided 3 times per week. Food was given to satiation (4% of the fish body weight) per feeding day. The test fish were not fed 24 h before test start.
Food: Sera Vipan; SERA GMBH, 52518 Heinsberg, Germany
The fish were not fed during the exposure.

Acclimatization was not necessary, because the quality of the dilution water was equivalent to the water used for holding.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

Total Hardness from the dilution water (Study day 0): 62

Test temperature:

21 - 25 °C, controlled at ± 1 °C

Minimum Temp.: 21.8 °C
Maximum Temp.: 22.2 °C
Mean Temp.: 21.9 °C

pH:

Control : 7,52 - 7,76
70 mg/L: 7,58 - 7,86

Dissolved oxygen:

Not less than 60% of air saturation value

Control : D1 100%; D1 90%; D2 87%
D2 after removal 96%; D3 85%; D4 81%
70 mg/L:D1 100%; D1 85%; D2 81%
D2 after removal 95%; D3 84%; D4 81%

Conductivity:

144 µS/cm (recent measurement 2020-04-27) holding of fish (same media like media used in the study)

Nominal and measured concentrations:

70 mg/L was tested as a threshold concentration in a limit test. The selection of the test concentration was based on the derivation of a threshold concentration (TC) from the data of the REACH dossier of the test item of results of a daphnia acute immobilization test (EC50 (0-48 h) = >100 mg/L) and of an alga toxicity test (ErC50 (0-72 h) = 70 mg/L).

Details on test conditions:

Test method:
A semi-static test with a renewal of the test media every two days was performed.

Limit concentration:
70 mg/L was tested as a threshold concentration in a limit test. The selection of the test concentration was based on the derivation of a threshold concentration (TC) from the data of the REACH dossier of the test item of results of a daphnia acute immobilization test (EC50 (0-48 h) = >100 mg/L) and of an alga toxicity test (ErC50 (0-72 h) = 70 mg/L).

Preparation of the test medium:
An appropriate amount of the test item was weighed and transferred into a test vessel filled with an appropriate amount of the dilution water. The test media was mixed with an ultraturrax (1 min, 17000 rpm). This procedure was repeated for every water renewal.

Control:
7 fish in dilution water (without test item but treated in the same way as the test solution) were tested under the same test conditions as the test replicates.

Reference item:
No reference item is recommended for this test according to the guidelines.

Duration of the test:
96 hours.

Test vessel:
Glass aquaria of 3 L were used (dimensions: 11.5/13/20 cm, depth of water: approx. 17 cm) and covered with a glass plate.

Test volume:
About 2.5 L per vessel

Aeration:
No aeration was provided.

Dilution water:
Same as used for holding

Acclimatization:
Acclimatization was not necessary, because the quality of the dilution water was equivalent to the water used for holding.

Number of fish:
7 zebrafish were used each for the limit concentration and the control. For the whole study 14 fish were used.

Loading, length of fish:
The fish density in the test vessels was 0.0924 g fish per liter test solution. Average body length at the test end: 1.74 cm
Average body weight at the test end: 0.033 g

Age of fish:
Juvenile

Replicates:
One replicate each for the limit concentration and the control

Introduction of fish:
The fish were randomly distributed among the treatments.

Feeding of test fish:
The fish were not fed during the exposure.

Water temperature (target):
21 - 25 °C, controlled at ± 1 °C

Dissolved oxygen Concentration (target):
Not less than 60% of air saturation value

Light intensity (target):
540 – 1000 Lux

Photoperiod:
A photoperiod (light / dark cycle: 16 / 8 h) was maintained during exposure.

The pH-value, temperature and oxygen saturation were measured in all test vessels at the start and the end of exposure as well as daily and after renewal of the test media from fresh and old test media.
Chlorine and nitrate were measured from each batch of dilution water used in the test.
Total hardness and TOC were determined at the start of the exposure in the dilution water. During the test, the water temperature was recorded continuously (once per hour) with a data logger. The light intensity on the surface of the test aquaria was measured at the start of the exposure.
The solution appearance and behavior was observed daily before and after renewal.

Reference substance (positive control):

not required

Remarks:

No reference item is recommended for this test according to the guidelines.

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

> 70 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.:

> 70 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The test item does not show toxic effects on fish at the test concentration of 70 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

The effect level is given based on the nominal test item concentration of Propan-1-ol, 2-amino-2-methyl-, 4-methylbenzenesulphonate. The threshold concentration of the test item at the nominal concentration 70 mg/L was found to have no lethal or toxic effects on zebrafish.
In conclusion, the LC0 corresponds to the nominal concentration of ≥ 70 mg test item/L, the LC50 was > 70 mg/L.

Executive summary:

The acute toxicity of the test item Propan-1-ol, 2-amino-2-methyl-, 4-methylbenzenesulphonate  to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (2019) from 2020-05-11 to 2020-05-15 at the test facility.
The test item is a white powder. A limit concentration of 70 mg/L was tested as a threshold concentration. The concentration was based on the derivation of a threshold concentration (TC) from the data of the REACH dossier of the test item of results of a daphnia acute immobilization test (EC50 (0-48 h) = >100 mg/L) and of an alga toxicity test (ErC50 (0-72 h) = 70 mg/L).
The study was performed over a period of 96 hours under semi-static conditions with renewal of test solutions after every 48 hours. Seven test organisms were exposed to the test concentration and the control, respectively. 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 test solutions were visually clear throughout the study.

The concentration of the test item Propan-1-ol, 2-amino-2-methyl-, 4-methylbenzenesulphonate was analytically verified via HPLC-DAD in the fresh media at the start of the exposure and at one renewal interval (0 hours and 48 hours) and in the corresponding 24-hours aged media after 48 and 96 hours in the limit concentration and the control. The measured concentration of the test item were 102% and 104% in the fresh media and 100% and 101% in the old media of the nominal test item concentration. Details of the analytical method are presented in section 11. The analytical results are presented in Table 6.
All validity criteria of the test guideline were met.  The test item does not show toxic effects on fish at the test concentration 70 mg/L.

Description of key information

The effect level is given based on the nominal test item concentration of Propan-1-ol, 2-amino-2-methyl-, 4-methylbenzenesulphonate. The threshold concentration of the test item at the nominal concentration 70 mg/L was found to have no lethal or toxic effects on zebrafish.
In conclusion, the LC0 corresponds to the nominal concentration of ≥ 70 mg test item/L, the LC50 was > 70 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Remarks:

no lethal or toxic effects on zebrafish at the limit concentration

Effect concentration:

> 70 mg/L