Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

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:
2018-11-13 to 2019-01-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD, EU Guideline Study: GLP
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
adopted July 17, 1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
May 30, 2008
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD No 23; Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures
Version / remarks:
December 15, 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
Name: TEMPIC
Synonym: Thiocure® TEMPIC
Chemical Name: Tris[2-(3-mercaptopropionyloxy)ethyl]isocyanurat
CAS No.: 36196-44-8
Batch No.: 0061488
Purity: 95.9 % according to certificate of analysis
Manufacturing Date: December 21, 2017
Certificate of Analysis Date: December 20, 2017
Aggregate State at Room Temperature: Liquid
Colour: Light yellow
Density: 1.30 - 1.36 g/cm3 according to MSDS
Expiry Date: December 20, 2018
Storage Conditions at Test Facility: At 4 +/- 4 °C, in the dark
Analytical monitoring:
yes
Remarks:
LC-MS/MS
Details on sampling:
Duplicate samples from the freshly prepared test media of all test concentrations and the control were taken at the start of the test and at the test medium renewal on day 3.
For the determination of the stability of the test item under the test conditions, respectively the maintenance of the test item concentrations during the test period, samples were taken in duplicate out of all test media and the control at the test medium renewal on day 1 and at the end of the test (96 hours) from the approximate centre of the aquaria.
However, from the test concentrations of 45.5 mg test item/L duplicate samples were taken on day 2 (aged test medium) because all fish were dead and no further samples were taken from this concentration. From the test concentration of 100 mg test item/L no samples were taken from day 3 and test end, because all fish of this concentration were dead before.
All samples were diluted by a factor of two with acetonitrile.
Additional samples of the control and of the dilution solvent were taken at each sampling without any sample treatment.
Vehicle:
no
Details on test solutions:
Test Concentrations: 100, 45.5, 20.7, 9.4 and 4.3 mg test item/L (spacing factor 2.2) and a control
corresponding to following mean measured concentrations of the test item: 23.4, 12.0, 5.05, 2.18 and 1.22 mg test item/L and a control. Control: In the control, test water was used without addition of the test item.
Dosage of Test Item:
The test item is not well soluble in test medium. To avoid physical effects of undissolved test item, no concentrations above the solubility limit of the test item in test water was tested. Therefore, a defined amount of the test item was added directly to the test water for the preparation of a stock solution and was carefully stirred for 24 hours in the dark to dissolve as much of the test item as possible. After cessation of mixing and a following period (1 hour) of settling to allow phase separation, the aqueous phase, i.e. the water accommodated fraction, was drawn off carefully and used as the highest test item concentration of 23.4 mg test item/L. Adequate volumes of this water accommodated fraction were mixed into test water to obtain the desired test item concentrations of 100, 45.5, 20.7, 9.4 and 4.3 mg test item/L (corresponding to mean measured concentrations of 23.4, 12.0, 5.05, 2.18 and 1.22 mg test item/L.) The test media were prepared just before introduction of the test fish (= start of the test and test medium renewal on day 1, 2 and 3).
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Juvenile Rainbow Trout (Oncorhynchus mykiss) mean length: 4.36 cm ± 0.3 cm; source: Forellenzuchtbetrieb Störk, 88348 Bad Saulgau, Germany
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
no
Hardness:
2.5 mmol/L (= 250.0 mg/L) as CaCO3
Test temperature:
14 - 15 °C
pH:
7.6 to 8.0
Dissolved oxygen:
87 to 99 %
Nominal and measured concentrations:
100, 45.5, 20.7, 9.4 and 4.3 mg test item/L (spacing factor 2.2) and a control
corresponding to following mean measured concentrations of the test item: 23.4, 12.0, 5.05, 2.18 and 1.22 mg test item/L and a control. Control: In the control, test water was used without addition of the test item.
Details on test conditions:
Test Environment: Controlled environmental room
Measurement of pH-Values, Dissolved Oxygen and Water Temperature: The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the freshly prepared and aged test media of each treatment group.
Water Temperature: 14 to 15 °C
pH-Values of the Test Water: 7.6 to 8.0
Dissolved Oxygen Concentrations: 92 to 101 % of the air saturation value
Light Regime: 16 h light : 8 h dark; 30 min dawn/dusk period was provided
Light Intensity: 560 to 760 lux Aeration of the Test Water: The test media were slightly aerated during the test.
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
5.05 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
12 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
7.79 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
In the control and at the concentrations up to 5.05 mg test item/L, all fish survived until the end of the experiment and no signs of intoxication occurred. At the test item concentrations of 12.0 and 23.4 mg test item/L, all fish died after 48, respectively after 24 hours of exposure.
Reported statistics and error estimates:
The LC50 and the corresponding confidence intervals were determined as geometric mean of the highest concentration showing 0 % (5.05 mg/L) mortality and the lowest concentration showing 100 % mortality (12.00 mg/L). The corresponding confidence intervals at 48, 72 and 96 h were determined as the tested concentrations surrounding the calculated EC50 values. The LC20 and LC10 could not be determined. The NOEC, the LOEC, the LC0 and the LC100 were determined directly from the raw data. The software used to perform the statistical analysis was ToxRat Professional, Version 3.2.1, ToxRat Solutions GmbH.
Validity criteria fulfilled:
yes
Remarks:
Control Mortality: In the control no fish died during the test. Dissolved Oxygen Concentration: The dissolved oxygen concentration did not fall below 92 % of air saturation value during the test.
Conclusions:
The toxic effect of the test item TEMPIC to Rainbow Trout (Oncorhynchus mykiss) was assessed in a semi-static concentration-response test. Based on the test results the 96-hour LC50 was determined to be 7.79 mg test item/L based on mean measured concentrations. The NOEC was determined to be 5.05 mg test item/L also based on mean measured concentrations. The initial concentrations and the maintenance of the exposure concentrations during the test were verified in the analytical part. All reported results refer to mean measured values since the concentrations of the test item were not within ± 20 % of the nominal concentrations during the test.
Executive summary:

Title: TEMPIC: Acute Toxicity to Rainbow Trout (Oncorhynchus mykiss) in a 96-hour Semi-Static Test

Purpose:

The purpose of this study was to evaluate the acute toxicity of the test item TEMPIC to fish. For this purpose, juvenile Rainbow Trout were exposed in a semi-static test to various concentrations under defined conditions for 96 hours. The recorded effects were the mortality and sublethal effects on the fish. The test method of application and the test system are recommended by the test guidelines and Rainbow Trout is one of the recommended test species. The purpose of the analytical part of this study was to verify the concentrations of the test item in the test medium.

Guidelines:

Commission Regulation (EC) No 440/2008, Annex, Part C, C.1.: "Acute Toxicity for Fish", Official Journal of the European Union, May 30, 2008

OECD Guideline for Testing of Chemicals, Section 2, No. 203, "Fish, Acute Toxicity Test", adopted July 17, 1992

OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000

SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

Materials and Methods

Test Item: TEMPIC; batch no.: 0061488; purity: 95.9 %, according to certificate of analysis

Test Species: Juvenile Rainbow Trout (Oncorhynchus mykiss) mean length: 4.36 cm ± 0.3 cm; source: Forellenzuchtbetrieb Störk, 88348 Bad Saulgau, Germany

Test Design: This study encompassed 6 treatment groups (5 dose rates of the test item and a control) each containing 7 individuals. The acute toxicity to unfed juvenile Rainbow Trout was determined in an aerated, semi-static, 96-hour test. The test fish were observed at test start and after approximately 2, 24, 48, 72 and 96 hours test duration for sublethal effects and mortality. Dead fish were removed at least once daily and discarded.

Endpoints: NOEC after 96 h, LOEC after 96 h; LC50: lethal concentration producing 50 % mortality after 96 h of exposure.

Test Concentrations:

100, 45.5, 20.7, 9.4 and 4.3 mg test item/L (spacing factor 2.2) and a control

corresponding to following mean measured concentrations of the test item: 23.4, 12.0, 5.05, 2.18 and 1.22 mg test item/L and a control.

Test Conditions:

Water temperature: 14 to 15 °C; pH value: 7.6 to 8.0; dissolved oxygen concentration: 92 to 101 % of the air saturation value; photoperiod: 16 h light - 8 h dark; light intensity: 560 to 760 lux and thus were within the ranges requested by guideline OECD 203.

Results:

Biological test results:

In the control and at the concentrations up to 5.05 mg test item/L, all fish survived until the end of the experiment and no signs of intoxication occurred. At the test item concentrations of 12.0 and 23.4 mg test item/L, all fish died after 48, respectively after 24 hours of exposure. Based on the test results the 96-hour LC50 was determined to be 7.79 mg test item/L based on mean measured concentrations. The NOEC was determined to be 5.05 mg test item/L also based on mean measured concentrations.

Analytical test results:

The quantification of the test item TEMPIC in the test samples was performed using liquid chromatography with MS/MS detection. In the freshly prepared test media at the start of the test and at the renewal of the test media 25 % of the nominal test concentrations were found (average of all test concentrations). In the aged test media after 24 or 96 hours test duration, 25 % of the nominal value was determined (average of all test concentrations available). During the test the test organism were exposed to a mean of 25 % of nominal.

Conclusion:

The toxic effect of the test item TEMPIC to Rainbow Trout (Oncorhynchus mykiss) was assessed in a semi-static concentration-response test. Based on the test results the 96-hour LC50 was determined to be 7.79 mg test item/L based on mean measured concentrations. The NOEC was determined to be 5.05 mg test item/L also based on mean measured concentrations. The initial concentrations and the maintenance of the exposure concentrations during the test were verified in the analytical part. All reported results refer to mean measured values since the concentrations of the test item were not within ± 20 % of the nominal concentrations during the test.

This study is classified acceptable and satisfies the guideline requirements for fish acute test studies.

Description of key information

OECD TG 203 study conducted with TEMPIC.

Key value for chemical safety assessment

LC50 for freshwater fish:
7.79 mg/L

Additional information

A NOEC = 5.05 mg/L could be reliable reported from this study.