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EC number: 218-485-4
CAS number: 2162-73-4
study was performed under GLP according to OECD TG 210. Due to the fast
hydrolysis of the test material (Neuland, 2020) the OECD Series on
Testing and Assessment, No. 23 "Guidance Document on Aqueous-phase
Aquatic Toxicity Testing of Difficult Test Chemicals" has also been
taken into account.
purpose of this study was to evaluate the toxicity of the registered
substance to the early-life stages of fish. For this purpose, fertilised
eggs of zebrafish (Danio rerio) were exposed in a flow-through
test to aqueous test media containing the test item at nominal
concentrations of 33 and 105µg/L under defined conditions. In parallel a
solvent control (100 µL DMF/L) and a water control were tested. Due to
the test item properties it was decided not to prepare the test media
constantly by using a constant flow rate by peristaltic pumps or syringe
pumps but to prepare the test media manually twice a day. In order to
keep the test design in a manageable dimension the experiment will be
running with two test concentrations only. According to OECD 210 this
extended limit design is acceptable for the determination of a NOEC.
several independent pre-experiments to determine the solubility and
stability of the test material in the test medium and the need to
develop a reproducible analytical method, dimethylformamide (DMF) was
used as solvent additive to achieve stable and reproducible
concentrations in the flow-through system.
test duration was 30 days after hatching. The eggs and larvae were
observed daily for survival, hatching, abnormal appearance and
behaviour. Additionally, at the end of the test, the individual length
of all surviving fish was determined using a digital caliper. Also, at
the end of the test all surviving fish were weighed (wet weight after
blotting dry and dry weight) in groups by test vessel (replicate). Dead
larvae were removed at least once daily and discarded. The test item
concentrations in the test water taken at test start and after DAI 7,
14, 21, 28 and 34 were chemically analysed by LC-MS/MS.
TRIDI is hydrolytically unstable with a t(1/2) of 37 seconds at a
temperature of 20 °C, it will be degraded during toxicity studies in
aqueous media. Based on the result of a hydrolysis study according to
OECD 111, 1,3,5-triisopropyl-2,4-diaminobenzene (TRIDA) will be formed
as major hydrolysis product. Therefore, the quantification of the
hydrolysis product 1,3,5-triisopropyl-2,4-diaminobenzene (TRIDA) was
performed using liquid chromatography with MS/MS detection and the
obtained results refer to the theoretical amount of TRIDA which can be
built from TRIDI considering equimolar conversion.
time weighted arithmetic mean measured concentrations of 27.5 and 95.8
µg test item/L were determined. However, as the mean test recoveries of
the nominal test concentration varied between 83 and 91% (all test
concentrations considered) the test item concentrations were determined
to be stable during the run of the test and results were based on
nominal test concentrations:
success: NOEC ≥ 105 µg test item/L
NOEC ≥ 105 µg test item/L
Length: NOEC ≥ 105 µg test item/L
Weight (wet): NOEC ≥ 105 µg test item/L
Weight (dry): NOEC ≥ 105 µg test item/L
Key_Long-term toxicity to fish: NOEC (30d) ≥
105 µg/L nominal (95.8 µg/L based on time weighted average
concentrations) for Danio rerio (flow-through, freshwater, OECD
Weight (dry): NOEC ≥ 105 µg test item/L
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