1-Butanone, 3-(dodecylthio)-1-[(1R,2S)-2,6,6-trimethyl-3-cyclohexen-1-yl]-, rel-

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 12 January to 03 March 2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

This study was performed according to OECD Guideline 203 and EU Method C1 with GLP statement. Based on the very low water solubility of the test substance, it's technically not feasible to obtain a relevant endpoint value.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

30th August 2005

Specific details on test material used for the study:

- Storage condition of test material: Approximately 4 °C under nitrogen in the dark

Analytical monitoring:

yes

Details on sampling:

See "Details on analytical methods"

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution of 1mg/ml in dimethylformamide (DMF) was made by serial dilutions in DMF. 2ml of this 1mg/ml stock solution were then dispersed into a volume of 20 l of dechlorinated tap water to give a 0.1 mg/l test concentration. This method of preparation was conducted in duplicate to give replicates R1 and R2. Each of the solvent stock solutions were inverted several times to ensure adequate mixing and homogeneity.
- Controls: The concentration and stability of the test material in the test centrifuged and and untreated samples were verified by chemical analysis at 0 (fresh media), 24, 48, 72 (old and fresh media) and 96 (old media) hours
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethylformamide (DMF)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Stock solutions: 100 % Final test solutions: 0.1 %
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: Oncorhynchus mykiss
- Source: Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK
- Age at study initiation (mean and range, SD): juvenile
- Length at study initiation (length definition, mean, range and SD): 4.8 cm (s.d. - 0.2) measured at the end of the study
- Weight at study initiation (mean and range, SD): 1.36 g (s.d. - 0.21) measured at the end of the study
- Loading rate: 0.48 g bodyweight/l
- Method of breeding: Fish were maintained in a glass fibre tank with a "single pass" water renewal system.
- Feeding during test: no


ACCLIMATION
- Acclimation period: 15 February to 27 February 2006 (13 days)
- Acclimation conditions (same as test or not): same
- Type and amount of food: commercial trout pellets (discontinued ca. 24 hours before start of the definitive test)
- Feeding frequency: not mentioned in study report

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not Applicable

Hardness:

100 mg/L CaCO3

Test temperature:

13-14°C (See Table 6.1.1/3 in "Any other information on results incl. Tables")

pH:

7.5 - 7.7 (See Table 6.1.1/3 in "Any other information on results incl. Tables")
No treatmeent related differences of pH

Dissolved oxygen:

10.2-10.5 mg O2/l (See Table 6.1.1/3 in "Any other information on results incl. Tables")

Salinity:

Not applicable

Nominal and measured concentrations:

See Tables 6.1.1/4 and 6.1.1/5 in "Any other information on results incl. Tables"

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 20 liters glass exposure vessels covered to reduce evaporation
- Type of flow-through (e.g. peristaltic or proportional diluter): Not Applicable (semi-static conditions)
- Renewal rate of test solution (frequency/flow rate): daily renewal of test medium
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.48 g /l


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water dechlorinated by passage through an activated carbon filter (Purite Series 500) and partly softened (Elga Nimbus 1248D Duplex Water Softener)
- Total organic carbon: 1.220-1.990 mg/l (Average:1.675 mg/l)
- Particulate matter: in the Precribed Concentration Value
- Metals: in the Precribed Concentration Value
- Pesticides: in the Precribed Concentration Value
- Chlorine: in the Precribed Concentration Value
- Alkalinity: in the Precribed Concentration Value
- Ca/mg ratio: in the Precribed Concentration Value
- Conductivity: 292.0 - 549.0 uS/cm @ 20°C (average: 437.3 uS/cm)


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h light / 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : behaviour, sub-lethal effects & death at 3, 6, 24, 48, 72 & 96 hours and pH, mg O2, T°C at 0, 24, 42, 72 & 96 hours (See Tables 6.1.1/1, 6.1.1/2 and 6.1.1/3 in "Any other information on results incl. Tables")


TEST CONCENTRATIONS
- Spacing factor for test concentrations: Not Applicable
- Justification for using less concentrations than requested by guideline: The test concentration of 0.1 mg/l was the highest attainable test concentration that could be prepared due to the limited solubility of the test material in water and having due regard to the amount of auxiliary solvent permitted in the study under the OECD Guidelines. At higher test concentrations there was a marked precipitation of the test material on addition of the solvent stock solution to water.
- Range finding study
- Test concentrations: 0.10 mg/l using static conditions
- Results used to determine the conditions for the definitive study: no mortalities were observed at 0.1 mg/l. Based on this information a single test concentration in duplicate of 0.1 mg/I was selected for the definitive study (See Table 6.1.1/1 in "Any other information on results incl. Tables")

Reference substance (positive control):

not required

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality & behaviour

Remarks on result:

other: 95% CL: -

Duration:

96 h

Dose descriptor:

other: Highest test concentration causing no mortality or sub-lethal effect

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality & behaviour

Remarks on result:

other: 95% CL: -

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 71 µg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

other: Mortality & behaviour

Remarks on result:

other: 95% CL: -

Duration:

96 h

Dose descriptor:

other: Highest test concentration causing no mortality or sub-lethal effect

Effect conc.:

71 µg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

other: Mortality & behaviour

Remarks on result:

other: 95% CL: -

Details on results:

See "Any other information on results incl. Tables" for more details

- Behavioural abnormalities: none
- Other biological observations: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none, test preparations were observed to be clear colorless solutions throughout the duration of the test.
- Effect concentrations exceeding solubility of substance in test medium: not applicable

Results with reference substance (positive control):

Not Applicable

Reported statistics and error estimates:

Not Applicable as one concentration was measured

Sublethal observations / clinical signs:

Table 6.1.1/1: Cumulative Mortality data

a.   Range-Finding test:

Concentration (mg/l)	Cumulative mortality (initial population : 3)
	3h	6h	24h	48h	72h	96h
Solvent Control	0	0	0	0	0	0
0.10	0	0	0	0	0	0

b.   Definitive test:

Concentration (mg/l)	Cumulative mortality (initial population : 7)						% mortality
	3h	6h	24h	48h	72h	96h	96h
Control	0	0	0	0	0	0	0
Solvent Control	0	0	0	0	0	0	0
0.10 R1	0	0	0	0	0	0	0
0.10 R2	0	0	0	0	0	0	0

 

Table 6.1.1/2: Behavioral Observations

Concentration (mg/l)	Abnormality	Time
		3h	6h	24h	48h	72h	96h
Control	No abnormalities detected	-	-	-	-	-	-
Solvent Control	No abnormalities detected	-	-	-	-	-	-
0.1 R1	No abnormalities detected	-	-	-	-	-	-
0.1 R2	No abnormalities detected	-	-	-	-	-	-
R1/R2: replicate 1 or replicate 2 - = nothing significant detected

 

Table 6.1.1/3: Environmental measurements

Concentration (mg/l)	0 hours (fresh media)			24 hours (old media)			24 hours (fresh media)
	pH	mg O2/l	T°C	pH	mg O2/l	T°C	pH	mg O2/l	T°C
Control	7.6	10.4	13.3	8.0	10.5	13.0	7.6	10.3	13.3
Solvent control	7.7	10.4	13.3	8.0	10.5	13.2	7.7	10.3	13.3
0.10 R1	7.7	10.5	13.4	8.0	10.5	13.0	7.7	10.4	13.4
0.10 R2	7.6	10.5	13.4	7.9	10.5	13.0	7.7	10.4	13.4
Concentration (mg/l)	48 hours (fresh media)			48 hours (old media)			72 hours (fresh media)
	pH	mg O2/l	T°C	pH	mg O2/l	T°C	pH	mg O2/l	T°C
Control	7.6	10.3	13.3	8.0	10.4	13.0	8.0	10.2	13.0
Solvent control	7.6	10.3	13.2	8.0	10.5	13.1	8.0	10.4	13.1
0.10 R1	7.6	10.3	13.3	8.0	10.5	13.0	8.0	10.3	13.0
0.10 R2	7.7	10.2	13.2	8.0	10.5	13.0	8.0	10.3	13.1
Concentration (mg/l)	72 hours (old media)			96 hours (old media)
	pH	mg O2/l	T°C	pH	mg O2/l	T°C
Control	7.6	10.5	13.2	8.0	10.5	13.0
Solvent control	7.7	10.4	13.2	8.0	10.5	13.0
0.10 R1	7.7	10.4	13.2	8.0	10.4	13.0
0.10 R2	7.6	10.4	13.2	7.9	10.4	13.1

Remark:100% of Air Saturation value (ASV) correspond to 10.5 mg 02/l this mean that all media had anASV > 96 %.

 

Table 6.1.1/4: Analysis of test sample solutions (Nominal vs measured concentrations)

Samples	Nominal concentration (mg/l)	Concentration found (mg/l)	Concentration expressed as % of nominal
0 hour (fresh)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	< LOQ 0.113 0.104 <LOQ 0.0956 0.102	- 113 104 - 96 102
24 hours (old)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	< LOQ 0.0927 0.0967 <LOQ 0.0774 0.0804	- 93 97 - 77 80
24 hours (fresh)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	< LOQ 0.100 0.118 <LOQ 0.0576/0.0135** 0.0672/0.0221**	- 100 118 -- 58/14** 67/22**
48 hours (old)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	< LOQ 0.0806 0.110 <LOQ 0.0555 0.0740	- 81 110 - 55 74
48 hours (fresh)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	< LOQ 0.116 0.118 <LOQ 0.0857 0.0947	- 116 118 - 86 95
72 hours (old)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	<LOQ 0.0875 0.0773 <LOQ 0.0598 0.0630	- 87 77 - 60 63
72 hours (fresh)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	<LOQ 0.122 0.127 <LOQ 0.0535/0.0144** 0.0643/0.00848**	- 122 127 - 54/14** 64/8**
96 hours (old)	Solvent Control U 0.10 R1 U 0.10 R2 U Solvent control C 0.10 R1 C 0.10 R2 C	<LOQ 0.0967 0.0965 <LOQ 0.0496 0.0601	- 97 97 - 50 60
LOQ: Limit ofQuantitation U = Untreated C = centrifuged at 40000g for 30 min at 21°C in polycarbonate tubes R1/R2: Replicates 1 and 2 ** Duplicate samples, stored and frozen prior to analysis

 

Chemical analysis of the freshly prepared test media at 0,24,48 and 72 hours showed measured concentrations to range from 100% to 127% of nominal for the untreated samples indicating that the test system was dosed correctly at each media renewal period.

Analysis of the centrifuged samples gave measured concentrations in the range 86% to 102% of nominal with the exception of the 24 and 72-Hour fresh centrifuged samples which showed measured concentrations ranging from 54% to 67%. These results were inconsistent with the results from analysis of the other centrifuged fresh media samples taken and hence were considered to be erroneous. Analysis of duplicate frozen samples showed measured concentrations of 8% to 22%. These results were also inconsistent with the results from analysis of the other centrifuged fresh media samples taken and hence were considered to also be erroneous as the corresponding centrifuged old or 'expired' test media samples taken following these dosing periods (at 48 and 96 hours) showed significantly higher measured test concentrations which were consistent with the centrifuged test sample results from the other dosing periods. The freezing and thawing of the duplicate samples was considered to have possibly degraded the test material hence the low measured concentrations.

Analysis of the 24-Hour old media samples at 24, 48, 72 and 96 hours showed measured concentrations to range from 77% to 110% of nominal for the untreated samples and from 50% to 80% of nominal for the centrifuged samples. The results obtained from the centrifuged test samples showed a decline in measured test concentration over the test period and wasinlinewith the results obtained from the media preparation trials conducted.

Given this decline in measured test concentrations it was considered justifiable to base the results on the time-weighted mean measured test concentrationsof the centrifuged test media to give a "worst case" analysis of the data. The time-weighted mean measured test concentrations were calculated as follows:

 

Table 6.1.1/5: Nominal and time-weighted mean measured concentrations

Nominal Concentration (mg/l)	Time-weighted Mean Measured Concentration (mg/l)	% of Nominal Concentration
0.10	0.071	71

Inspection of the immobilization data gave the following data:

Table 6.1.1/6: LC50 values

	Mean measured concentrations
Time (h)	LC50 (mg/l)	95% C.L.
3	> 0.071	-
6	> 0.071	-
24	> 0.071	-
48	> 0.071	-
72	> 0.071	-
96	> 0.071	-
Highest test concentration causing no effects	0.071mg/l

 

 

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test material to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LC50 of greater than 0.10 mg/L.
Based on the time-weighted mean measured test concentrations of the centrifuged test media the acute toxicity of the test material to rainbow trout gave a 96-Hour LC50 value of greater than 0.071 mg/L. The highest test concentration causing no mortality or sub-lethal effect was 0.071 mg/L.

Executive summary:

Introduction.A study was performed to assess the acute toxicity of the test material to rainbow trout(Oncorhynchusmykiss).The method followed that described in the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish,AcuteToxicity Test" referenced as Method C.1 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC.

 

Methods.A Determination of the GeneralPhysico-chemical Properties study conducted on the test material (SafepharrnLaboratories Limited Project Number: 161/439) showed the water solubility value of the test material to be 0.090 mg/l using the shake flask method of preparation.

Preliminary solubility work showed that the highest attainable test concentration (by visual inspection of the test media) was 0.10 mg/l using a preliminary solution prepared indimethylformamide. Media preparation trials were conducted in order to determine the amount of dissolved test material present and the most appropriate method of test media preparation. The results of the media preparation trials indicated that using a solvent spike method of preparation was the most suitable for this test material and indicated that after centrifugation at 40000 g for 30 minutes a dissolved concentration of approximately 0.055 mg/l was obtained. Given the range of tests to be conducted on this material in order to maintain consistency it was considered inappropriate to centrifuge the test preparations prior to exposure. Instead samples were taken for analysis following centrifugation, the results of which gave an indication of the dissolved and hence bioavailable test concentration.

Following a preliminary range-finding test fish were exposed, in two groups of seven, to an aqueous solution of the test material, at a single concentration of 0.1 mg/l for a period of 96 hours at a temperature of approximately 14°C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

 

Results.The 96-Hour LC50 based on nominal test concentrations was greater than 0.10 mg/L. The test concentration of 0.1 mg/L was the highest attainable test concentration due to the limited solubility of the test material in water and auxiliary solvent, and having due regard for the amount of auxiliary solvent permitted in the test under the OECD Guidelines.

 

Chemical analysis of the freshly prepared test media at 0, 24, 48 and 72 hours showed measured concentrations to range from 100% to 127% of nominal for the untreated samples indicating that the test system was dosed correctly at each media renewal period. Analysis of the centrifuged samples gave measured concentrations in the range 86% to 102% of nominal with the exception of the 24 and 72-hour fresh centrifuged samples which showed measured concentrations of 54% to 67%. These results were inconsistent with the results from analysis of the other centrifuged fresh media samples taken and hence were considered to be erroneous. Analysis of duplicate frozen samples showed measured concentrations of 8% to 22%. These results were also inconsistent with the results from analysis of the other centrifuged fresh media samples taken and hence were considered to also be erroneous as the corresponding centrifuged old or 'expired' test media samples taken following these dosing periods (at 48 and 96 hours) showed significantly higher measured test concentrations which were consistent with the centrifuged test sample results from the other dosing periods. The freezing and thawing of the duplicate samples was considered to have possibly degraded the test material hence the low measured concentrations.

 

Analysis of the 24-hour old media samples at 24, 48, 72 and 96 hours showed measured concentrations to range from 77% to 110% of nominal for the untreated samples and from 50% to 80% of nominal for the centrifuged samples.The results obtained from the centrifuged test samples showed a decline in measured test concentration over the test period and was in line with the results obtained from the media preparation trials conducted.

Given this decline in measured test concentrations it was considered justifiable to base the results on the time-weighted mean measured test concentrations centrifuged test media to give a "worst case" analysis of the data.

 

The 96-Hour LC50 based on the time-weighted mean measured test concentrations of the centrifuged test media was greater than 0.071 mg/L. The highest test concentration causing no mortality or sub-lethal effect was 0.071 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

From May 21, 2007 to June 28, 2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

This study was performed according to international guidelines without GLP statement. Based on the very low water solubility of the test substance, it's technically not feasible to obtain a relevant endpoint value.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: HJ/T153-2004

Deviations:

not specified

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

none

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.5 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

other: Highest concentration causing no mortality or effects

Effect conc.:

>= 0.5 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

In the controls and the treatments at the loading rate of 100 mg/L (mean measured concentration 0.5 mg/L) no fish death was determined during the limit test period of 96 hours.

The pH values in the treatments and the controls ranged from 7.69 to 8.24, and water temperature was maintained at 23°C±1°C. During the period of the test, the dissolved oxygen concentration was always 70% or higher (see table below).

Table 6.1.1/1: Periodical observation results in the controls and the treatments

		Controls		Treatments (100 mg/L)
Mean measured concentration (mg/L)		-		0.5
0h	Temp (°C)	23.9	23.9	23.9	23.9
	DO (% ASV)	93.7	93.5	93.2	93.4
	pH	8.14	8.15	8.24	8.22
	Death number	0	0	0	0
	Cumulative mortality (%)	0	0	0	0
24h	Temp (°C)	23.4	23.4	23.4	23.4
	DO (% ASV)	95.6	95.5	94.6	94.5
	pH	8.15	8.16	8.20	8.21
	Death number	0	0	0	0
	Cumulative mortality (%)	0	0	0	0
48h	Temp (°C)	23.1	23.1	23.1	23.1
	DO (% ASV)	95.7	96.2	91.5	91.2
	pH	8.15	8.16	8.22	8.22
	Death number	0	0	0	0
	Cumulative mortality (%)	0	0	0	0
72h	Temp (°C)	23.6	23.6	23.6	23.6
	DO (% ASV)	95.7	95.8	92.7	92.8
	pH	8.10	8.09	8.16	8.15
	Death number	0	0	0	0
	Cumulative mortality (%)	0	0	0	0
96h	Temp (°C)	23.5	23.5	23.5	23.5
	DO (% ASV)	75.9	78.0	77.3	74.1
	pH	7.69	7.71	7.80	7.85
	Death number	0	0	0	0
	Cumulative mortality (%)	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

Based on these results, the test substance has no acute toxicity effect on zebra fish up to its solubility limit in the water under the present conditions of the test, and its hazardous level is slight.

Executive summary:

The acute toxicity effect of the test substance to zebra fish (Brachydanio rerio) was determined in a 96-hour semi-static test with a daily test medium renewal, according to OECD Guideline n°203 and HJ/T153 -2004 without GLP statement. A limit test was performed in accordance with the result of the preliminary test to demonstrate that the test substaoce at a loading rate of 100 mg/L has no acute toxicity effect on the test organisms. Thus, only the treatments at a loading rate of 100 mg/L and the controls were tested.

Due to the low water solubility of the test substance and shortage of appropriate auxiliary solvents or dispersants, a supersaturated dispersion of the test substance with a loading rate of 100 mg/L was homogenized by continuously stirred over 4 hours prior to each test medium preparation. Then the dispersion was filtered with the 0.45 µm membrane filter. Only the undiluted filtrate with the maximum concentration of dissolved test substance was used as the test medium.

The analytically measured test substance concentrations in the analyzed test medium samples were 0.5mg/L and 0.4mg/L at the beginning and the end of the test respectively. The biological results are related to the treatments of 100 mg/L and to the mean measured test substance concentration of 0.5 mg/L (calculated as the average of the test concentration).

In the controls and the treatments at the loading rate of 100 mg/L (mean measured concentration 0.5 mg/L) no fish death was determined during the limit test period of 96 hours. Therefore, the highest concentration causing no mortality or effects to zebra fish were determined to be the loading rate of 100 mg/L (mean measured concentration 0.5 mg/L). The 96h-LC50 was clearly higher than the loading rate of 100 mg/L (mean measured concentration 0.5 mg/L).

In conclusion, the test substance had no acute toxic effects on zebra fish up to its solubility limit in test water under the present conditions of the test. Based on these results, the hazardous level of the test substance is slight.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Adequacy of study:

supporting study

Study period:

Run on 2013-03

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Remarks:

(Q)SAR method with established scientific validity. The substance falls within the applicability domain of the model.

Justification for type of information:

1. SOFTWARE
ECOSAR

2. MODEL (incl. version number)
ECOSAR v1.11

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
C/C=C/C(C1C(C)C=CCC1(C)C)=O
log Kow : 3.6 (estimated result based on differents QSARs)

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See "Any other information on results incl. tables"

5. APPLICABILITY DOMAIN
See "Any other information on results incl. tables"

6. ADEQUACY OF THE RESULT
See "Any other information on results incl. tables"

Qualifier:

according to guideline

Guideline:

other: REACH guidance on QSARs R.6, May 2008

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

(not relevant)

Test organisms (species):

no data

Test type:

not specified

Water media type:

not specified

Total exposure duration:

96 h

Remarks on exposure duration:

none

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.776 mg/L

Remarks on result:

other: ECOSAR Class: Neutral Organic SAR (Baseline toxicity)

Sublethal observations / clinical signs:

Validity of model:

1. Defined Endpoint: Acute toxicity to fish (lethality).

2. Unambigous algorithm: The ECOSAR class determined was Vinyl/Allyl Ketones. However, based on observed effects for fish, this chemical class is not expressing excess toxicity compared to neutral organics. The class-specific SAR equation is yielding lower toxicity values than estimated from baseline toxicity (neutral organics), therefore the neutral organics SAR is used to estimate the toxicity of the substancefor fish: Linear regression QSAR; Log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108. To convert the LC50 from mmol/L to mg/L, multiply by the molecular weight.

3. Applicability domain: applicable to chemicals with log Kow less than 5.0 and molecular weight less than 1000 g/mol. The model can be suitable for chemicals class of neutral organic.

4. Statistical characteristics: N = 296+55 and the coefficient of determination R² = 0.878.

5. Mechanistic interpretation: related to the partitioning of the substance from water into the organisms lipid phase.

Adequacy of prediction: the substance falls within the applicability domain described above and therefore the predicted value can be considered reliable.

Validity criteria fulfilled:

yes

Conclusions:

delta-Damascone was predicted to have an 96h-LC50 of 5.776 mg/L to fish after 96h exposure. The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Executive summary:

The acute toxicity to fish of delta-Damascone was estimated using the ECOSAR v1.11 QSAR model available from the U.S. EPA. The estimation 96h-LC50 was 5.776 mg/L (neutral organics class), calculated from an estimated log Kow value at 3.6 (mean of QSAR results) and a linear regression:

log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108.

The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Adequacy of study:

supporting study

Study period:

Run on 2013-03

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Remarks:

(Q)SAR method with established scientific validity. This substance is not related to an existing ECOSAR class. Estimation provided use the Neutral Organics QSAR equations which represent baseline toxicity potential (minimum toxicity) assuming a simple non-polar narcosis model. Without empirical data on structurally similar chemicals, it is uncertain if this substance will present significantly higher toxicity above baseline estimates. The substance falls within the applicability domain of the model.

Justification for type of information:

1. SOFTWARE
ECOSAR

2. MODEL (incl. version number)
ECOSAR v1.11

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
CC1(C)CC=CC(C)C1C(CC(SCC(O)=O)C)=O
log Kow : 3.1 (estimated result based on differents QSARs)

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See "Any other information on results incl. tables"

5. APPLICABILITY DOMAIN
See "Any other information on results incl. tables"

6. ADEQUACY OF THE RESULT
See "Any other information on results incl. tables"

Qualifier:

according to guideline

Guideline:

other: REACH guidance on QSARs R.6, May 2008

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

(not relevant)

Test organisms (species):

no data

Test type:

not specified

Water media type:

not specified

Total exposure duration:

96 h

Remarks on exposure duration:

none

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

240.24 mg/L

Remarks on result:

other: > limit of solubility. ECOSAR Class: Neutral Organics-acid (acid moeity found: predicted values multiplied by 10).

Sublethal observations / clinical signs:

Validity of model:

1. Defined Endpoint: Acute toxicity to fish (lethality).

2. Unambigous algorithm: This substance is not related to an existing ECOSAR class. Estimation provided use the Neutral Organics QSAR equations which represent baseline toxicity potential (minimum toxicity) assuming a simple non-polar narcosis model. Without empirical data on structurally similar chemicals, it is uncertain if this substance will present significantly higher toxicity above baseline estimates. Acid moeity was found, therefore predicted values were multiplied by 10: Linear regression QSAR; Log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108. To convert the LC50 from mmol/L to mg/L, multiply by the molecular weight.

3. Applicability domain: applicable to chemicals with log Kow less than 5.0 and molecular weight less than 1000 g/mol. The model can be suitable for chemicals class of neutral organic.

4. Statistical characteristics: N = 296+55 and the coefficient of determination R² = 0.878.

5. Mechanistic interpretation: related to the partitioning of the substance from water into the organisms lipid phase.

Adequacy of prediction: the substance falls within the applicability domain described above and therefore the predicted value can be considered reliable.

Validity criteria fulfilled:

yes

Conclusions:

delta-Damascone mercapto acetic acid was predicted to have an 96h-LC50 > solubility limit, estimated at 84.53 mg/L to fish after 96h exposure. The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Executive summary:

The acute toxicity to fish of delta-Damascone mercapto acetic acid was estimated using the ECOSAR v1.11 QSAR model available from the U.S. EPA. The estimation 96h-LC50 was greater than the solubility limit, estimated at 84.53 mg/L (neutral organics-acid), calculated from an estimated log Kow value at 3.1 (mean of QSAR results) and a linear regression (acid moeity was found, therefore predicted values were multiplied by 10):

log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108.

The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

(Q)SAR

Adequacy of study:

supporting study

Study period:

Run on 2013-03

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification

Remarks:

(Q)SAR method with established scientific validity. This substance is not related to an existing ECOSAR class. Estimation provided use the Neutral Organics QSAR equations which represent baseline toxicity potential (minimum toxicity) assuming a simple non-polar narcosis model. Without empirical data on structurally similar chemicals, it is uncertain if this substance will present significantly higher toxicity above baseline estimates. The substance falls within the applicability domain of the model.

Justification for type of information:

1. SOFTWARE
ECOSAR

2. MODEL (incl. version number)
ECOSAR v1.11

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
CC1(C)CC=CC(C)C1C(CC(SCCC(O)=O)C)=O
log Kow : 3.3 (estimated result based on differents QSARs)

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
See "Any other information on results incl. tables"

5. APPLICABILITY DOMAIN
See "Any other information on results incl. tables"

6. ADEQUACY OF THE RESULT
See "Any other information on results incl. tables"

Qualifier:

according to guideline

Guideline:

other: REACH guidance on QSARs R.6, May 2008

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Remarks:

(not relevant)

Test organisms (species):

no data

Test type:

not specified

Water media type:

not specified

Total exposure duration:

96 h

Remarks on exposure duration:

none

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

166.699 mg/L

Remarks on result:

other: > limit of solubility. ECOSAR Class: Neutral Organics-acid (acid moeity found: predicted values multiplied by 10).

Sublethal observations / clinical signs:

Validity of model:

1. Defined Endpoint: Acute toxicity to fish (lethality).

2. Unambigous algorithm: This substance is not related to an existing ECOSAR class. Estimation provided use the Neutral Organics QSAR equations which represent baseline toxicity potential (minimum toxicity) assuming a simple non-polar narcosis model. Without empirical data on structurally similar chemicals, it is uncertain if this substance will present significantly higher toxicity above baseline estimates. Acid moeity was found, therefore predicted values were multiplied by 10: Linear regression QSAR; Log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108. To convert the LC50 from mmol/L to mg/L, multiply by the molecular weight.

3. Applicability domain: applicable to chemicals with log Kow less than 5.0 and molecular weight less than 1000 g/mol. The model can be suitable for chemicals class of neutral organic.

4. Statistical characteristics: N = 296+55 and the coefficient of determination R² = 0.878.

5. Mechanistic interpretation: related to the partitioning of the substance from water into the organisms lipid phase.

Adequacy of prediction: the substance falls within the applicability domain described above and therefore the predicted value can be considered reliable.

Validity criteria fulfilled:

yes

Conclusions:

delta-Damascone mercapto propionic acid was predicted to have an 96h-LC50 > solubility limit, estimated at 47.31 mg/L to fish after 96h exposure. The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Executive summary:

The acute toxicity to fish of delta-Damascone mercapto propionic acid was estimated using the ECOSAR v1.11 QSAR model available from the U.S. EPA. The estimation 96h-LC50 was greater than the solubility limit, estimated at 47.31 mg/L (neutral organics-acid), calculated from an estimated log Kow value at 3.3 (mean of QSAR results) and a linear regression (acid moeity was found, therefore predicted values were multiplied by 10):

log 96h-LC50 (mmol/L) = -0.8981*log Kow + 1.7108.

The substance falls within the applicability domain and therefore the predicted value can be considered reliable.

Description of key information

Based on the very low water solubility of the registered substance, it's technically not feasible to obtain a relevant endpoint value. The substance had no acute toxic effects on fish up to and significantly higher than its solubility limit in test water.

Additional information

Two studies are available to asses the acute toxicity of the registered substance to fish (Oncorhynchus mykiss and Danio rerio) in a 96h semi-static test.

The first study (Safepharm, 2006), evaluated as the key study, was performed according to OECD Guideline No 203, EU Method C.1 with GLP statement and the second study (SAES, 2007), evaluated as the supporting study, was performed according to OECD Guideline No 203, HJ/T153 -2004 but without GLP statement.

Based on the very low water solubility of the substance, it's technically not feasible to obtain a relevant endpoint value. The substance had no acute toxic effects on fish up to and significantly higher than its solubility limit in test water.

Moreover, as a strong and consistent evidence of mineralisation of the registered substance was provided, the acute toxicities of the degradation products, which are the relevant species for the environmental risk assessment, were determined using the ECOSAR v1.11 QSAR model available from the U.S. EPA.. The assessed degradation products were Delta-Damascone mercapto acetic acid (DD-ma) (which is the main degradation product) and two other larget metabolites (Delta-Damascone mercapto proprionic acid (DD-mp) and Delta-Damascone (DD)). The estimated 96h-LC50 values were > 84.53 mg/L for DD-ma, > 47.31 mg/L for DD-mp and 5.78 mg/L for DD. According to these data, the degradation products are not expected to be toxic (T) in a PBT assessment context.