2-Butanone, 1,3,4-trihydroxy-

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:

2003-07-28 to 2003-08-01

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:

adopted 17 July 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

Version / remarks:

second edition, 15 July 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

December 1992

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Name: (S)-1,3,4-trihydroxybutan-2-one
- CAS no.: 533-50-6

Analytical monitoring:

yes

Details on sampling:

During the combined limit/range-finding test duplicate samples for analysis were taken from the blank control and 100 mg/L.

- Frequency: at t = 0, t = 24 h and t = 96 h
- Volume: 10 mL from approximate centre of the test vessels
- Storage: samples taken at t = 0 and t = 24 h were stored in a freezer until analysis
- Reserve samples: 10 mL taken for possible analysis from the blank control and 100 mg/L at t = 24 and t = 96 hours.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: preparation of stock solutions in test medium at 100 mg/L; no special treatment other than a magnetic stirring period of ca. 20 minutes was necessary to completely dissolve the test substance in the test medium; lower test concentrations were prepared by subsequent dilution of the stock in test medium
- Controls: negative control (test medium only)
- Test concentration separation factor: A combined limit/range-finding test was carried out. That means, the limit concentration of 100 mg/L was already included in the concentrations used for the range-finding study. For the range-finding a separation factor of 10 between nominal test item concentrations of 0.1, 1.0, 10 and 100 mg/L was applied. For the limit test a nominal concentration of 100 mg/L was considered.
- Evidence of undissolved material: no, test substance was completely soluble in test medium at tested concentrations and all final test solutions were clear and colourless

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Strain: not reported
- Source: Zodiac, proefacc, “De Haar Vissen”, L.U. Wageningen, The Netherlands
- Length at study initiation: mean length measured to be 2.36 +/- 0.32 cm
- Weight at study initiation: mean weight measured to be 0.41 +/- 0.17 g
- Characteristics: F1 from a single parent-pair bred

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Medium: ISO-medium, formulated using Milli-Ro water (tap water pourified by reverse osmosis; Millipore Corp., Bedford, Mass., USA)
- Type and amount of food during acclimation: Trouvit
- Feeding frequency during acclimation: daily
- Health during acclimation: mortality below 5 %

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

250 mg CaCO3/L (hardness of test medium)

Test temperature:

20.5 - 21.4 °C in negative control and 20.5 - 21.3 °C in treatment (limit concentration of 100 mg test item/L)

pH:

7.3 - 8.0 (negative control) and 7.3 - 7.8 (100 mg/L)

Dissolved oxygen:

6.4 - 8.7 mg/L (negative control) and 6.4 - 8.6 mg/L (100 mg/L)

Salinity:

not applicable

Conductivity:

not specified

Nominal and measured concentrations:

nominal: 0 (negative control), 100 mg/L (limit concentration); 0 (negative control), 98.55 mg/L (measured mean, n = 2, after 0 hours), 99.5 mg/L (measured mean, n = 2, after 24 hours), 97.9 mg/L (measured mean, n = 2, after 96 hours), corresponding to 97 - 101 % of nominal test item concentration over a period of 96 hours

Details on test conditions:

TEST SYSTEM
- Test vessel: 6.5 litres, all-glass
- Aeration: aeration introduced after 24 hours of exposure
- No. of organisms per vessel: blank control and 100 mg/L: 7 fish per test group; 0.1, 1.0 and 10 mg/L (range-finding): 3 fish per test group
- No. of vessels per concentration: 1
- No. of vessels per control: 1 for negative control; 1 per treatment in positive control (using 0.06, 0.10, 0.15, 0.22 and 0.32 mg pentachlorophenol/L in ISO medium)
- Biomass loading rate: blank control and 100 mg/L: 0.57 g fish/litre, i.e. 7 fish per 5 litres of test medium; 0.1, 1.0 and 10 mg/L: 0.25 g fish/litre, i.e. 3 fish per 5 litres of test medium

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium, aerated until the dissolved oxygen concentration had reached saturation and the pH had stabilised. After aeration the hardness was 250 mg CaCO3 per litre and the pH was 8.0.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light, 8 hours darkness
- Light intensity: not specified

EFFECT PARAMETERS MEASURED:
- Mortality and other effects: at 2 ¼, 24, 48, 72 and 96 hours following the start of exposure; dead fish removed when observed
- Fish length and weight: ten fish of the batch used for the test weighed prior to the start of the test
- Dissolved oxygen content, temperature and pH: daily in all vessels, beginning at the start of the test (day 0)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable to main test (limit test)
- Range finding study: yes, combined range-finding/limit test
- Test concentrations: ; 0.1, 1.0, 10 and 100 mg/L, whereby 100 mg/L was also considered as limit concentration
- Results used to determine the conditions for the definitive study: the definitive study was integrated in the range-finding study (combined range-finding/limit test)

Reference substance (positive control):

yes

Remarks:

reference substance: pentachlorophenol

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other abnormalities: no
- Observations on body length and weight: no
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no


ACCEPTABILITY OF THE STUDY
1. No abnormal behaviour (including mortality) was observed in the blank control.
2. The oxygen concentration was maintained at least at 60 % of the air saturation value throughout the test (> 5 mg/L at 22 °C). Other test conditions (pH and temperature) were maintained within the limits prescribed by the protocol and guidelines.
3. The analytical program provided clear evidence that the actual measured concentration was maintained at more than 80 % relative to the initial measured concentration during the test period.
4. The 96h-LC50 of the reference chemical for the stock of fish was in reasonable agreement with the results obtained previously in the laboratory.

Results with reference substance (positive control):

96h-LC50 = 0.21 mg/L (95 % confidence interval: 0.19 - 0.27 mg/L)

Reported statistics and error estimates:

No LC50 could be calculated because the test substance proved to be non-toxic (LC50 > max. tested concentration)

Sublethal observations / clinical signs:

Table 1: Incidence of mortality and total mortality during the combined limit/range-finding test

Concentration test item (mg/L)	Initial number of fish	Cumulative mortality					Total mortality (%)
		2.25 h	24 h	48 h	72 h	96 h
Blank-conrol	7	0	0	0	0	0	0
0.1	3	0	0	0	0	0	0
1	3	0	0	0	0	0	0
10	3	0	0	0	0	0	0
100	7	0	0	0	0	0	0

 

Table 2: Concentrations of test item in test medium (final test)

Time of sampling [hours]	Concentration
	Nominal [mg/L]	Analysed [mg/L]	Relative to nominal [%]	Relative to initial [%]
0	0	n.d. ^1,2	n.a.
	0	n.d. ^1,2	n.a.
	100	98.7 ^1,2	99
	100	98.4 ^1,2	98
24	0	n.d. ^1,2	n.a.	n.a.
	0	n.d. ^1,2	n.a.	n.a.
	100	101 ^1,2	101	102
	100	97.9 ^1,2	98	99
96	0	n.d. ^1,2	n.a.	n.a.
	0	n.d. ^1,2	n.a.	n.a.
	100	97.4 ^1,2	97	99
	100	98.4 ^1,2	98	100

1: Samples were combined with range-finding study

2: Samples were frozen until analysis

n.d.: not detected

n.a.: not applicable

Validity criteria fulfilled:

yes

Conclusions:

In an Acute Toxicity Study according to ISO International Standard 7346-1, EEC directive 92/69, Part C.1 and OECD TG 202 with Carp (Cyprinus carpio) the 96h-LC50 of the test item was determined to be > 100 mg/L (nominal).

Executive summary:

An Acute Toxicity Study with carp (Cyprinus carpio) was carried out in accordance with ISO International Standard 7346-1, EEC directive 92/69, Part C.1 and OECD TG 202. A limit test was combined with a range-finding test. Seven carp were exposed to a blank-control and a test item concentration of 100 mg/L for the limit test, while three carp per test group were exposed to 0.1, 1.0 and 10 mg/L for the combined range-finding test.  Samples for analytical confirmation of actual exposure concentrations were taken at the start, after 24 hours and at the end of the 96-hour test period. Analysis of the samples taken during the combined limit/range-finding test showed that the measured concentration was 97 – 101 % of the nominal test item concentration. The validity of the test was evaluated based on the validity criteria of current OECD TG 203 (dated 18 June 2019). In the blank-control the mortality did clearly not exceed 10% (no mortality observed). Further, the dissolved oxygen concentration was ≥60 % of the air saturation value in all test vessels throughout the exposure. Moreover, the nominal test item concentrations were analytically verified. Therefore, the study is regarded valid and acceptable for assessment.

Description of key information

In an Acute Toxicity Study according to ISO International Standard 7346-1, EEC directive 92/69, Part C.1 and OECD TG 202 with Carp (Cyprinus carpio) the 96h-LC50 of the test item was determined to be > 100 mg/L (nominal).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 100 mg/L