Reaction mass of 3-O-acetyl-1,5-anhydro-4-butyl-2,4-dideoxy-2-methyl-D-xylitol and (2R,3S)-2-methyloctane-1,3-diyl diacetate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: read-across from a guideline study

Justification for type of information:

The read-across justification is presented in the Endpoint summary Aquatic toxicity, because for fish, Daphnia, and algae the same substance is used, and the reasoning is included in one document. The accompanying files are also attached there.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

32 mg/L

Basis for effect:

mortality (fish)

Remarks on result:

other: read-across from Jasmal

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The range-finding test was conducted between 08 and 12 June 2015 and the final test between 20 and 24 July 2015.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

This information is used for read-across to Jessemal.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

(1992)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

(2008)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test concentrations were verified by chemical analysis. Water samples (15 mL from the approximate centre of the test vessels) were taken from the control and each exposure level at the start of the test and after 72 hours from the freshly prepared solutions, and at the first renewal (t=24) and the end of the test from the 24-h old solutions.
In addition, samples were taken from the 46 mg/L test solution at 48 hours of exposure when all fish in this concentration were found dead. Samples were stored in the freezer until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Jasmal was completely soluble in test medium at the concentrations tested.
* test medium: adjusted ISO medium
* start concentration: 100 mg/L
* stirring period: 44-85 minutes of magnetic stirring in a vessel to reach the maximum solubility of the test substance in the test medium
* test concentration range: lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp (Cyprinus carpio)
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Age at study initiation (mean and range, SD): not reported; F1 from a single parent-pair bred in UV-treated water
- Length at study initiation (final test): 2.3 ± 0.10 cm
- Weight at study initiation (final test): 0.23 ± 0.07 g
- Method of breeding: F1 from a single parent-pair bred in UV-treated water.
- Feeding during test: No feeding from 24 hours prior to the test and during the total test period.

ACCLIMATION
- Acclimation period: at least 12 days after delivery
- Acclimation conditions: same as test, i.e. adjusted ISO medium
- Type of food: pelleted fish food (Cyprico Crumble Excellent (300-500 µm))
- Feeding frequency: daily
- Health during acclimation: <5% mortality observed during the seven days prior to the start of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

180 mg CaCO3/L

Test temperature:

21.6 - 22.7°C

pH:

7.4 - 7.9

Dissolved oxygen:

6.1-9.4 mgO2/L

Nominal and measured concentrations:

Based on the results of the range-finding test (biological effects observed between 10 and 100 mg/L) the following nominal test concentrations were assigned to the final test: 4.6, 10, 22, 46 and 100 mg/L.

Measured test concentrations: see tables 1 and 2 in field 'Any other information on results'

The effect concentrations are based on nominal concentrations since the average measured concentrations derived from the 24 h intervals were within 80-99% of nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: 3 litres, all-glass
- Type: open
- Fill volume: 3 litres of test solution
- Aeration: no
- Renewal rate of test solution: 24 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.54 g fish/litre, i.e.7 fish per 7 litres of test medium

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium with a hardness of 180 mg CaCO3 per litre and a pH of 7.7 ± 0.3

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Mortality (after 2¼, 24, 48, 72 and 96 hours following the start of exposure)
- Other effects: clinical effects (such as hypoactive swimming, loss of equilibrium or immobility) - daily

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.1- 2.2

- Range finding study
- Test concentrations: 0.1, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

pentachlorophenol (January 2015)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Nominal concentrations were analytically confirmed (average exposure concentrations were 80-99% of nominal). 95% CL 22-46 mg/L

Details on results:

The effect, i.e. 50% mortality, was reached already after 48 hours of exposure.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: 100% at 0.46 mg/L
- LC50: 0.32 mg/L
- Other: effect (i.e. 50% mortality) reached already within 48 h of exposure

Reported statistics and error estimates:

LC50 determination
* For the responses recorded at 24 hour of exposure: the maximum likelihood estimation method with the weibits of the percentages of dead fish as function of the logarithms of the corresponding concentrations was used. Calculations were performed with ToxRat Professional 3.0.0. (ToxRat Solutions®, GmbH, Germany).
* For the remaining time points: The LC50 could not be determined using the maximum likelihood estimation method with the weibits of the percentages of dead fish as function of the logarithms of the corresponding concentrations. 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)½, with A and B being limits of the 95% confidence interval.

Sublethal observations / clinical signs:

Table 1: Concentrations of the test item in test medium - final test

Time of sampling [hours]	Concentration		Relative to nominal [%]	Relative to initial [%]
	Nominal [mg/L]	Analysed [mg/L]
0	0	n.d.	n.a.
	4.6	4.35	95
	10	9.79	98
	22	21.7	99
	46	46.0	100
	100	98.7	99
24 (old)	0	n.d.	n.a.	n.a.
	4.6	3.72	81	86
	10	8.22	82	84
	22	17.5	79	80
	46	40.6	88	88
48 (old)	46	43.9	95	n.a.
72 (fresh)	0	n.d.	n.a.
	4.6	3.74	81
	10	9.79	98
	22	19.0	87
96 (old)	0	n.d.	n.a.	n.a.
	4.6	2.98	65	80
	10	7.56	76	77
	22	14.5	66	76

n.d.     Not detected.

n.a.     Not applicable.

Table 2: Average exposure concentrations vs. nominal concentrations

Jasmal Nominal concentration (mg/L)	Measured concentration (mg/L)					Average exposure concentration (mg/L)
	Day 0 (fresh)	Day 1 (old)	Day 2 (old)	Day 3 (fresh)	Day 4 (old)
4.6	4.4	3.7	n.d.	3.7	3.0	3.7 (80)
10	9.8	8.2	n.d.	9.8	7.6	8.8 (88)
22	22	18	n.d.	19	15	18 (82)
46	46	41	44	n.d.	n.d.	43* (93)
100	99	n.d.	n.d.	n.d.	n.d.	99 (99)

n.d. – not determined

* - calculated based on results obtained in the first refreshment

() – between brackets the percentage of nominal are given

Table 3: Incidence of mortality and total mortality during the final test

Jasmal Nominal concentration (mg/L)	Initial number of fish	Cumulative mortality					Total Mortality (%)
		2¼h	24h	48h	72h	96h
Control	7	0	0	0	0	0	0
4.6	7	0	0	0	0	0	0
10	7	0	0	0	0	0	0
22	7	0	0	0	0	0	0
46	7	0	2	7	7	7	100
100	7	7	7	7	7	7	100

Table 4: Clinical effects observed during the final test

Jasmal Nominal concentration (mg/L)	Time of recording (hours)	Specification of effects	Relative number
22	24-72	Hypoactive swimming	7/7
46	2¼	Immobile Hypoactive and loss of equilibrium	3/7 4/7
	24	Immobile	5/5

Validity criteria fulfilled:

yes

Remarks:

no mortality in controls; test conditions were maintained constant throughout the test; dissolved O2 concentration was at least 60% of the air saturation value throughout the test; concentration of the substance was maintained within 20% of nominal

Conclusions:

The 96 h LC50 value in fish (Cyprinus carpio) is 32 mg/L.

Executive summary:

A study was performed to assess the acute toxicity of the substance to fish (Cyprinus carpio). The study was conducted in accordance with OECD Guideline for Testing of Chemicals No. 203. In a semi-static test set-up in open 3-L vessels seven fish per concentration were exposed for 96 hours to 5 concentrations of substance in test water (nominal concentrations up to 100 mg/L). Throughout the test, test solutions were refreshed in intervals of 24 hours. Clinical effects (such as hypoactive swimming, loss of equilibrium or immobility) and incidences of mortality were recorded for the control and each test group at t=2, 24, 48 and 72 hours and the following values were determined: 96 h LC50 = 32 mg/L. All results are expressed in terms of nominal concentrations. Measured concentrations ranged from 81 - 100 % of nominal in fresh solutions, but decreased to 76 - 88 % of nominal during the refreshment period of 24 hours. Therefore, average exposure concentrations were calculated, however, the mean of these average concentrations was found to be within 80 - 99 % of nominal.

Description of key information

The short-term toxicity of Jessemal to fish is assessed based on read-across from the close structural analogue Jasmal (CAS# 18871-14-2), resulting in an LC50 fish value of 32 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

32 mg/L

Additional information

The read across rationale is presented in the Aquatic Endpoint summary because also Daphnia and algae toxicity are based on read across and these are addressed together. The accompanying files are also attached there. Below the executive summary of Jasmal fish toxicity is presented.

Jasmal's fish toxicity

A study was performed to assess the acute toxicity of the substance to fish (Cyprinus carpio). The study was conducted in accordance with OECD Guideline for Testing of Chemicals No. 203. In a semi-static test set-up in open 3-L vessels seven fish per concentration were exposed for 96 hours to 5 concentrations of substance in test water (nominal concentrations up to 100 mg/L). Throughout the test, test solutions were refreshed in intervals of 24 hours. Clinical effects (such as hypoactive swimming, loss of equilibrium or immobility) and incidences of mortality were recorded for the control and each test group at t=2, 24, 48 and 72 hours and the following values were determined: 96 h LC50 = 32 mg/L. All results are expressed in terms of nominal concentrations. Measured concentrations ranged from 81 - 100 % of nominal in fresh solutions, but decreased to 76 - 88 % of nominal during the refreshment period of 24 hours. Therefore, average exposure concentrations were calculated, however, the mean of these average concentrations was found to be within 80 - 99 % of nominal.