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Ecotoxicological information

Short-term toxicity to fish

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short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: according to EC Directive 92/69/EEC and Regulation EC/440/2008 guideline methods under GLP conditions
according to guideline
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
Specific details on test material used for the study:
Batch No 11029
Purity 98,3%
white solified mass
Analytical monitoring:
Details on sampling:
Analytical measurements in the pre-test showed that the test substance is stable during the 96 hours exposure period. Test was performed as a static test, samples were taken at the beginning and the end of the exposure period. 10 ml were taken from the centre of the test vessel
Details on test solutions:
Test substance, a solidified mass, was heted for 1 hour at 90°C until it became a clear yellow viscious liquid. The test substance was completely soluble in the test medium at 100 mg/L. Magnetic stirring was applied to dissolve the test substance in the test medium. All other concentrations were prepared by dilution. Final test solutions were clear and colourless.
Test organisms (species):
Cyprinus carpio
Details on test organisms:
Source of fish: Zodiac, proefacc, "De Haar Vissen", L.U., Wageningen, the Netherlands
Mean length Range-finding test: 3.02 +/- 0.23 cm, Final test: 2.7 +/-0.24 cm
Mean weight Range-finding test: 1.12 +/- 0.30 g, Final test: 0.48 +/- 0.1 8 g
Characteristics: F1 from a single parent-pair bred in UV-treated water.
Reason for selection: This system has been selected as an internationally
accepted species.
Total fish used: 54
QuarantineIAcclimation At least 12 days after delivery
In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.
Test type:
Water media type:
Total exposure duration:
96 h
250 mg CaCO3/l
Test temperature:
The test temperature ranged from 20.2 to 20.8 degrees C. 
The pH ranged from 8.1 to 7.5 
Dissolved oxygen:
dissolved oxygen ranged from 5.9 to 9.2 mg/l
Nominal and measured concentrations:
nominal concentrations final test: 10, 18, 32, 56 and 100 mg/L
measured concentration final test 0 hours (concentrations of 10, 32 and 100 mg/L were measured): 9.51, 28.3 and 88.6 mg/L
measured concentration final test 96 hours (concentrations of 10 and 32 mg/L were measured): 8.48 and 27.3 mg/L
Details on test conditions:
Test type Static
Test vessels 6 litres, all-glass.
Test medium: ISO-medium, aerated until the dissolved oxygen
concentration had reached saturation and the pH had stabilised. After aeration the hardness was 250 mg CaC03 per litre and the pH was 8.0.
Number of fish: 7 fish per concentration and control
Loading: 0.67 g fishflitre, i.e. 7 fish per 5 litres of test medium.
Illumination: 16 hours photoperiod daily
Aeration: Aeration was introduced after 24 hours of exposure
Feeding: No feeding from 48 hours prior to the test and during the total test period.
Introduction of fish: Directly after preparation of the test media
96 h
Dose descriptor:
Effect conc.:
10 mg/L
96 h
Dose descriptor:
Effect conc.:
32 mg/L
Key result
96 h
Dose descriptor:
Effect conc.:
42 mg/L
96 h
Dose descriptor:
Effect conc.:
56 mg/L
Results with reference substance (positive control):
96 hours LC50 with pentachlorophenol was 0.09 mg/L
Reported statistics and error estimates:
The LC50 could not be determined using the maximum likelihood estimation method with the
probits of the percentages of dead fish as function of the logarithms of the corresponding
concentrations (Finney, D.J., 1971 : Probit analysis, Cambridge University Press, Cambridge,
U.K., 3rd edition). This was because there was no concentration between the highest
concentration (A) at which 0% mortality and the lowest concentration (B) at which 100%
mortality occurred. Instead, the LC50 was calculated as (AB)" , with A and B being limits of the
95% confidence interval
Sublethal observations / clinical signs:

The 24h- and 96h-LC50 for carp exposed to 2,4,7,9-Tetramethyl-5-Decyne-4,7-Diol was 42 mg/l with 0
percent mortality at 32 mg/l (LC0) and 100 percent mortality at 56 mg/l (LC100). Concentrations down to 18 mg/l induced effects on swimming behavior and pigmentation, while no sub-lethal effects occurred at 10 mg/l.

Validity criteria fulfilled:
The 96h-LC50 for carp exposed to 2,4,7,9-TETRAMETHYL-5-DECYNE-4,7-DIOL was already
reached after 24 hours of exposure and was 42 mg/l with 0% mortality at 32 mg/l (LC0) and
100% mortality at 56 mg/l (LC100). Concentrations down to 18 mg/l induced effects on swimming
behaviour and pigmentation, while no sub-lethal effects occurred at 10 mg/l.
Executive summary:

First a range-finding test was conducted with exposure of
fish to 0.1, 1.0, 10 and 100 mg/l. All fish died at 100 mg/l
within 2.5 hours after the start of the test, while no fish
died at 10 mg/l. Analysis of samples taken at 10 mg/l showed
that this concentration was stable during the 96-hour test

After the range-finding test, a final test was performed
with carp exposed to concentrations of 0, 10, 18, 32, 56 and
100 mg/l in a static system. Seven carp were exposed per
concentration and a control. The loading was 0.67 g
fish/liter.  At the start of test, the water hardness was
250 mg CaCO3 per liter, the dissolved oxygen concentration
was saturated, and the water pH was 8.0.  The dissolved
oxygen, water pH, and temperature were measured daily. 
Samples for analytical confirmation of actual exposure
concentrations were taken at the start and the end of the

Analysis of the samples taken during the final test showed
that the measured concentrations in the samples taken at the
start of the test were 9.5 mg/l (95 percent), 28.3 mg/l (89
percent) and 88.6 mg/l (89 percent) at the nominal
concentrations of 10, 32 and 100 mg/l, respectively. The
measured concentrations at 32 and 100 mg/l could be related
to lower recoveries found at 180 mg/l in a recovery control
experiment (87-90 percent). Hence, the actual concentrations
in these samples were considered to be in agreement with
nominal.  At the end of the test period measured
concentrations had not decreased by more than 20 percent of
the initial concentrations. As a result, toxicity parameters
were based on nominal concentrations.

Description of key information

The 96h-LC50 for carp exposed to 2,4,7,9-TETRAMETHYL-5-DECYNE-4,7-DIOL was already
reached after 24 hours of exposure and was 42 mg/l with 0% mortality at 32 mg/l (LC0) and
100% mortality at 56 mg/l (LC100). Concentrations down to 18 mg/l induced effects on swimming
behaviour and pigmentation, while no sub-lethal effects occurred at 10 mg/l

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Dose descriptor:
Effect concentration:
42 mg/L

Additional information