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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was conducted between 19 June 2011 and 7 July 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Principles of method if other than guideline:
In view of the difficulties associated with the evaluation of aquatic toxicity of poorly water soluble test items, a modification of the standard method for the preparation of aqueous media was performed. An approach endorsed by several important regulatory authorities in the EU and elsewhere (ECETOC 1996, OECD 2000 and Singer et al 2000), is to expose organisms to a Water Accommodated Fraction (WAF) of the test item in cases where the test item is a complex mixture and is poorly soluble in water and in the permitted auxiliary solvents and surfactants. Using this approach, aqueous media are prepared by mixing the test item with water for a prolonged period. At the completion of mixing, the test item phase is separated by siphon and the test organisms exposed to the aqueous phase or WAF (which may contain dissolved test item and/or leachates from the test item). Exposures are expressed in terms of the original concentration of test item in water at the start of the mixing period (loading rate) irrespective of the actual concentration of test item in the WAF.
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Single nominal loading rate of 100 mg/l.The concentration and stability of the test item in the test preparations were verified by chemical analysis at 0 (fresh media), 24 and 96 hours (old media) (see details on analytical methods section).- Sampling method: Water samples were taken from the control and each replicate test vessel at 0 (fresh media), 24 and 96 hours (old media) for quantitative analysis. The samples taken at 0 and 24 hours were stored at approximately -20°C prior to analysis. Duplicate samples and samples at 24 (fresh media), 48 and 72 hours (fresh and old media) were taken and stored at approximately -20°C for further analysis, if necessary. The method of analysis, recovery and test preparation analyses are described in details on analytical methods section.
Vehicle:
no
Details on test solutions:
VALIDATION OF MIXING PERIOD:Pre-study investigational work was carried out to determine whether stirring for a prolonged period produced significantly higher measured test concentrations in the WAF.A WAF of nominal loading rate of 100 mg/l was prepared, in duplicate, in deionised reverse osmosis purified water. One loading rate was stirred for a period of 23 hours and the other for a period of 71 hours. After a 1-Hour standing period the mixtures were then removed by siphon and the concentration of the test item in the 100 mg/l loading rate WAF was verified by chemical analysis (see Appendix 3: Validation of Mixing Period - any other information on results incl tables section).RANGE-FINDING TEST:Due to the low aqueous solubility and complex nature of the test item, for the purposes of the range-finding test the test item was prepared as a Water Accommodated Fraction (WAF).The loading rate to be used in the definitive test was determined by a preliminary range-finding test.In the range-finding test fish were exposed to a nominal loading rate of 100 mg/l. A single loading rate was used as results of the range-finding test for the Acute Toxicity to Daphnia magna study (Harlan Laboratories Ltd., Project Number: 41101227) indicated that toxicity was not expected at this loading rate.An amount of test item (2100 mg) was added to the surface of 21 litres of dechlorinated tap water to give the 100 mg/l loading rate. After addition of the test item the dechlorinated tap water was stirred using a magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixture allowed to stand for 1 hour. Microscopic observations made on the WAFs indicated that no microscopic particles of test item or micro-dispersions were present in the water column. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and the WAF removed by mid-depth siphoning (the first approximate 75-100 ml discarded) to give the 100 mg/l loading rate WAF.In the range-finding test 3 fish were added to each 20 litre test and control vessel and maintained at approximately 14°C in a temperature controlled room with a photoperiod of16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours under static test conditions.The control group was maintained under identical conditions but not exposed to the test item.Each vessel was covered to reduce evaporation. After 3, 6, 24, 48, 72 and 96 hours any mortalities or sub-lethal effects of exposure were determined by visual inspection of the test fish.A sample of the WAF preparation was taken for chemical analysis at 0 hours (fresh media) and 24 hours (old media) in order for determine the stability of the test item under test conditions. All samples were stored at approximately -20°C prior to analysis.DEFINITIVE TEST:Based on the results of the range-finding test a "Limit test" was conducted at a single loading rate of 100 mg/l to confirm that no mortalities or sub-lethal effects of exposure were observed.EXPERIMENTAL PREPARATION:Due to the low aqueous solubility and complex nature of the test item for the purposes of the definitive test the test item was prepared as a Water Accommodated Fraction (WAF).An amount of test item (2100 mg) was added to the surface of 21 litres of dechlorinated tap water to give the 100 mg/l loading rate. After addition of the test item the dechlorinated tap water was stirred using a magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixture allowed to stand for 1 hour. Microscopic observations made on the WAFs indicated that no microscopic particles of test item or micro-dispersions were present in the water column. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and the WAF removed by mid-depth siphoning (the first approximate 75-100 ml discarded) to give the 100 mg/l loading rate WAF.This method of preparation was conducted in duplicate to give replicates R1 and R2 .The concentration and stability of the test item in the test preparations were verified by chemical analysis at 0 (fresh media), 24 and 96 hours (old media) (see attached background material Appendix 4: Chemical Analysis of Test Loading Rates).
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
The test was carried out using juvenile rainbow trout (Oncorhynchus mykiss). Fish were obtained from Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK and maintained in house since 5 May 2011. Fish were maintained in a glass fibre tank with a "single pass" water renewal system. Fish were acclimatised to test conditions from 15 June 2011 to 27 June 2011. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods.The water temperature was controlled at approximately 14°C with a dissolved oxygen content of greater than or equal to 8.6 mg O2/l. These parameters were recorded daily. The stock fish were fed commercial trout pellets which was discontinued approximately 24 hours prior to the start of the definitive test. There was zero mortality in the 7 days prior to the start of the test and the fish had a mean standard length of 4.4 cm (sd = 0.3) and a mean weight of 1.23 g (sd = 0.22) at the end of the definitive test. Based on the mean weight value this gave a loading rate of 0.43 g bodyweight/litre.The diet and diluent water are considered not to contain any contaminant that would affect the integrity and outcome of the study.Test WaterThe test water used for both the range-finding and definitive tests was the same as that used to maintain the stock fish.Laboratory tap water was dechlorinated by passage through an activated carbon filter (Purite Series 500) and partly softened (Elga Nimbus 1248D Duplex Water Softener) giving water with a total hardness of approximately 140 mg/l as CaCO3. After dechlorination and softening the water was passed through a series of computer controlled plate heat exchangers to achieve the required temperature. Typical water quality characteristics for the tap water as supplied, prior to dechlorination and softening, are given in Appendix 2: Typical Water Quality Characteristics (see attached background material).
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
Any mortalities and sub-lethal effects of exposure were recorded at 3, 6, 24, 48, 72 and 96 hours after the start of exposure. The criteria of death were taken to be the absence of both respiratory movement and response to physical stimulation.
Hardness:
Total hardness of approximately 140 mg/l as CaCO3.
Test temperature:
The test vessels were covered and maintained at approximately 14ºC
pH:
The pH was measured using a Hach HQ30d Flexi Handheld meter.pH range of control ranged from 7.1-7.9 from measurements taken at 0, 24, 48, 72 and 96 hours with fresh and old media.pH range of test solutions ranged from 7.0 - 7.7 from measurements taken at 0, 24, 48, 72 and 96 hours with fresh and old media.Please see Physico-Chemical Measurements appendix 5 see in any other information on results incl. tables section.
Dissolved oxygen:
The dissolved oxygen concentration was measured using a Hach HQ30d Flexi Handheld meter..Please see Physico-Chemical Measurements appendix 5 (see in any other information on results incl. tables section).
Salinity:
Freshwater used.
Nominal and measured concentrations:
Based on the results of the range-finding test a "Limit test" was conducted at a single (nominal) loading rate of 100 mg/l to confirm that no mortalities or sub-lethal effects of exposure were observed.
Details on test conditions:
Exposure conditions:As in the range-finding test 20 litre glass exposure vessels were used for each test concentration. At the start of the test seven fish were placed in each test vessel at random, in the test preparations. The test vessels were then covered to reduce evaporation and maintained at approximately 14ºC in a temperature controlled room with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours. The test vessels were aerated via narrow bore glass tubes. The fish were not individually identified and received no food during exposure.The control group was maintained under identical conditions but not exposed to the test item.A semi-static test regime was employed in the test involving a daily renewal of the test preparations to ensure that the concentrations of the test item remained near nominal and to prevent the build up of nitrogenous waste products.Any mortalities and sub-lethal effects of exposure were recorded at 3, 6, 24, 48, 72 and 96 hours after the start of exposure. The criteria of death were taken to be the absence of both respiratory movement and response to physical stimulation.Physico-chemical measurements:The water temperature, pH and dissolved oxygen concentrations were recorded daily throughout the test. The measurements at 0 hours, and after each test media renewal at 24, 48 and 72 hours, represent those of the freshly prepared test preparations while the measurements taken prior to each test media renewal, and on termination of the test after 96 hours, represent those of the used or 24-Hour old test preparations. The pH and the dissolved oxygen concentration were measured using a Hach HQ30d Flexi Handheld meter and the temperature was measured using Hanna Instruments HI 93510 digital thermometer.
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limits not stated
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95 % CL not stated
Details on results:
Range-finding Test:Cumulative mortality data from the exposure of rainbow trout to the test item during the range-finding test are given in Table 1 (see in any other information on results incl. tables section). There were no sub-lethal effects of exposure during the range-finding test.The results showed no mortalities at 100 mg/l loading rate WAF.Chemical analysis of the 100 mg/l loading rate WAF test preparation at 0 and 24 hours (see Appendix 4: Chemical Analysis of Test Loading Rates, in attached background material) showed measured test concentrations of 94 and 95 mg/l were obtained respectively thereby indicating that the test item was stable under test conditions.Based on this information, a single loading rate, in duplicate, of 100 mg/l using a stirring period of 23 hours followed by a 1-Hour standing period was selected for the definitive test. This experimental design conforms to a "Limit test" to confirm that no mortalities or sub-lethal effects of exposure were observed.Definitive Test:Mortality data:Cumulative mortality data from the exposure of rainbow trout to the test item during the definitive test are given in Table 2 (see in any other information on results incl. tables section).There were no mortalities in 14 fish exposed to a 100 mg/l loading rate WAF for a period of 96 hours. Inspection of the mortality data gave the following results:Time (h)LL*50 (mg/l)3>1006>10024>10048>10072>10096>100The No Observed Effect Loading rate (NOEL) was considered to be 100 mg/l loading rate WAF. The No Observed Effect Loading rate is based upon zero mortalities and the absence of any sub-lethal effects of exposure at this loading rate.It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/l. Sub-lethal effects:There were no sub-lethal effects of exposure observed in 14 fish exposed to a 100 mg/l loading rate WAF for a period of 96 hours.Physico-chemical measurements:The results of the physico-chemical measurements are given in Appendix 5 (see any other information on results incl tables section). Temperature was maintained at approximately 14ºC throughout the test, while there were no treatment related differences for oxygen concentration or pH.Vortex depth measurements:The vortex depth was recorded at the start and end of each mixing period and was observed to be a dimple at the water surface on each occasion (see Table 3 see in any other information on results incl. tables section). Observations on test item solubility:Observations on the test media were carried out during the mixing and testing of the WAF.At the start of the mixing period the 100 mg/l loading rate WAF was observed to be a clear, colourless water column with oily globules of test item at the surface of the water, bottom of the vessel and dispersed throughout. At the end of the mixing period and following the 1-Hour standing period the 100 mg/l loading rate WAF was observed to be a clear, colourless water column with an oily layer of test item on the surface. Microscopic inspection of the WAF showed no microscopic particles or microdispersions of test item to be present. During the test the 100 mg/l loading rate was observed to be a clear, colourless solution.Chemical analysis of test loading rates:Information supplied by the Sponsor, indicated that the test item contained approximately 42% 2-ethylhexyl dihydrogen phosphate (MW = 210 g/mol) and 49% bis(2-ethylhexyl) hydrogen phosphate (MW = 322 g/mol)). Under the acidic conditions of the analysis, both compounds could be observed, but the lower acidity and greater response of the bis(2-ethylhexyl) hydrogen phosphate meant that it appeared as the main peak in the chromatogram.Analysis of the test preparations (see Appendix 4 in attached background material) at 0 (fresh media), 24 and 96 hours (old media) showed measured concentrations to range from 94.9 to 98.0 mg/l with the exception of the 100 mg/l loading rate WAF replicate R2 at 0 and 24 hours which showed measured concentrations of 60.7 and 66.0 mg/l respectively. Analysis of the duplicate samples, stored at approximately -20°C prior to analysis, showed measured concentrations of 69.7 and 68.9 mg/l respectively. The results were lower than expected as results from the remainder of the samples showed measured concentrations in excess of 90 mg/l. A review of the data could not reveal a reason for this, however, this anomalous result was considered to have no adverse effect on the outcome of the test.Given that toxicity cannot be attributed to a single component or a mixture of components but to the test item as a whole the results were based on nominal loading rates only.
Results with reference substance (positive control):
No positive control.

Table 1              Cumulative Mortality Data in the Range-finding Test

Nominal

Loading Rate

(mg/l)

Cumulative Mortality

(Initial Population = 3)

3 Hours

6 Hours

24 Hours

48 Hours

72 Hours

96 Hours

Control

0

0

0

0

0

0

100

0

0

0

0

0

0

Table 2              Cumulative Mortality Data in the Definitive Test

Nominal

Loading Rate

(mg/l)

Cumulative Mortality
(Initial Population =7)

%

Mortality

3 Hours

6 Hours

24 Hours

48 Hours

72 Hours

96 Hours

96 Hours

Control

0

0

0

0

0

0

0

100 R1

0

0

0

0

0

0

0

100 R2

0

0

0

0

0

0

0


R1and R2= Replicates 1 and 2

Table 3              Vortex Depth Measurements at the Start and End of Each Mixing Period

FIRST MIXING PERIOD

 

Nominal Loading Rate (mg/l)

Control

100 R1

100 R2

*

+

*

+

*

+

Height of Water Column (cm)

36.0

36.0

36.0

36.0

36.0

36.0

Depth of Vortex (cm)

~0.2

~0.2

~0.2

~0.2

~0.2

~0.2

Observation of Vortex

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

SECOND MIXING PERIOD

 

Nominal Loading Rate (mg/l)

Control

100 R1

100 R2

*

+

*

+

*

+

Height of Water Column (cm)

36.0

36.0

35.9

35.9

35.9

35.9

Depth of Vortex (cm)

~0.2

~0.2

~0.2

~0.2

~0.2

~0.2

Observation of Vortex

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

THIRD MIXING PERIOD

 

Nominal Loading Rate (mg/l)

Control

100 R1

100 R2

*

+

*

+

*

+

Height of Water Column (cm)

36.0

36.0

35.9

35.9

36.0

36.0

Depth of Vortex (cm)

~0.2

~0.2

~0.2

~0.2

~0.2

~0.2

Observation of Vortex

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

FOURTH MIXING PERIOD

 

Nominal Loading Rate (mg/l)

Control

100 R1

100 R2

*

+

*

+

*

+

Height of Water Column (cm)

36.0

36.0

36.0

36.0

36.0

36.0

Depth of Vortex (cm)

~0.2

~0.2

~0.2

~0.2

~0.2

~0.2

Observation of Vortex

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present

Dimple present


R1- R2= Replicates 1 and 2

*= Start of mixing period

+= End of mixing period

Appendix 3:      Validation of Mixing Period

Pre-study investigational work was carried out to determine whether stirring for a prolonged period produced significantly higher measured levels of test item, in the WAF. A WAF of a nominal loading rate of 100 mg/l was prepared in duplicate in deionised reverse osmosis purified water and stirred using a stirring rate such that a vortex was formed to give a dimple at the water surface. One loading rate was stirred for a period of 23 hours and the other for a period of 71 hours. After a 1-Hour standing period the mixtures were then removed by siphon and samples taken for chemical analysis. 

The results are summarised as follows:

Test Item Loading Rate

(mg/l)

Measured Concentration (mg/l)

Time (hours)

24

72

Control

<LOQ

<LOQ

100

59.3

82.9

Although the results from this pre-study investigational work showed that the measured concentration increased from 24 hours to 72 hours, chemical analysis of the 100 mg/l Loading rate WAF test samples from the range-finding test showed that 24 hours was a sufficient preparation period to ensure the maximum amount of the dissolved water soluble fraction of the test item. The preparation of the WAF was maintained at 24 hours.


 LOQ = Limit of Quantitation (assessed down to 0.03 mg/l)

Appendix 5      Physico-Chemical Measurements

Nominal

Loading Rate

(mg/l)

Time (Hours)

0 Hours (Fresh Media)

24 Hours (Old Media)

24 Hours (Fresh Media)

pH

mg O2/l

%ASV*

TºC

pH

mg O2/l

%ASV*

T°C

pH

mg O2/l

%ASV*

T°C

Control

7.1

9.7

96

15

7.9

10.1

98

14

7.1

9.8

97

15

100 R1

7.2

9.7

96

15

7.8

9.8

95

14

7.1

9.7

96

15

100 R2

7.1

9.7

96

15

7.8

9.8

95

14

7.2

9.7

96

15

 

Nominal

Loading Rate

(mg/l)

Time (Hours)

48 Hours (Old Media)

48 Hours (Fresh Media)

72 Hours (Old Media)

pH

mg O2/l

%ASV*

TºC

pH

mg O2/l

%ASV*

T°C

pH

mg O2/l

%ASV*

T°C

Control

7.3

10.0

97

14

7.2

10.1

100

15

7.1

10.2

99

14

100 R1

7.5

9.6

93

14

7.1

9.9

99

15

7.5

9.9

96

14

100 R2

7.5

9.7

94

14

7.0

9.8

98

15

7.6

9.9

96

14

 

Nominal

Loading Rate

(mg/l)

Time (Hours)

72 Hours (Fresh Media)

96 Hours (Old Media)

pH

mg O2/l

%ASV*

TºC

pH

mg O2/l

%ASV*

T°C

Control

7.7

10.1

100

15

7.4

10.4

101

14

100 R1

7.6

9.9

98

15

7.5

9.9

96

14

100 R2

7.4

9.8

97

15

7.7

9.9

96

14


*ASV= Dissolved oxygen concentration expressed as a percentage of Air Saturation Value

R1and R2= Replicates 1 and 2

 

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LL50 value of greater than 100 mg/l loading rate WAF. Correspondingly the No Observed Effect Loading rate was 100 mg/l loading rate WAF.The test was considered to be valid as all validation criteria were met.
Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Regulation (EC) No. 440/2008.

Methods

Following a preliminary range-finding test, fish were exposed, in groups of seven, to a Water Accommodated Fraction (WAF) of the test item, at a single nominal loading rate of 100 mg/l for a period of 96 hours at a temperature of approximately 14ºC 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 LL50based on nominal loading rates was greater than 100 mg/l loading rate WAF and correspondingly the No Observed Effect Loading rate was 100 mg/l loading rate WAF.

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/l.

Information supplied by the Sponsor, indicated that the test item contained approximately 42% 2-ethylhexyl dihydrogen phosphate (MW = 210 g/mol) and 49% bis(2-ethylhexyl) hydrogen phosphate (MW = 322 g/mol)).   Under the acidic conditions of the analysis, both compounds could be observed, but the lower acidity and greater response of the bis(2-ethylhexyl) hydrogen phosphate meant that it appeared as the main peak in the chromatogram.

Analysis of the test preparations at 0 (fresh media), 24 and 96 hours (old media) showed measured concentrations to range from 94.9 to 98.0 mg/l with the exception of the 100 mg/l loading rate WAF replicate R2 at 0 and 24 hours which showed measured concentrations of 60.7 and 66.0 mg/l respectively. Analysis of the duplicate samples, stored at approximately -20°C prior to analysis, showed measured concentrations of 69.7 and 68.9 mg/l respectively. The results were lower than expected as results from the remainder of the samples showed measured concentrations in excess of 90 mg/l. A review of the data could not reveal a reason for this, however, this anomalous result was considered to have no adverse effect on the outcome of the test.

Given that toxicity cannot be attributed to a single component or a mixture of components but to the test item as a whole the results were based on nominal loading rates only.


Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across hypothesis proposed is that the organism is not exposed to common compounds but rather, because of structural similarity, that different compounds have similar ecotoxicological and fate properties. In this case the ECHA Read-Across Assessment Framework (RAAF) Scenario 2 is used.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Primary Source: Phosphoric acid, C8-12 (even-numbered) alkyl esters [EC not yet assigned; CAS not assigned]
Supporting Source: Phosphoric acid, 2-ethylhexyl ester [EC 235-741-0; CAS 12645-31-7]
Target: Phosphoric Acid, linear and branched Octyl Esters [EC not yet assigned; CAS not assigned]
3. ANALOGUE APPROACH JUSTIFICATION
Available data for the Primary Source substance suggest that algae are the most sensitive species. This is confirmed by data available for Fish, Daphnia and algae from studies performed on the Supporting Source substance. In the available studies the effects levels are similar in both species. In this case, it is acceptable to read-across for the short-term fish toxicity endpoint from the Supporting Source to the Target.
4. DATA MATRIX
Short-term toxicity to fish:
Primary Source: No data
Supporting Source: 96-h LL50 value of greater than 100 mg/L loading rate WAF
Target: No data
Short-term toxicity to aquatic invertebrates:
Primary Source : Daphnia magna; OECD TG 202.
48-h EC50 > 5.62 mg/L (measured) (100 mg/L nominal loading rate WAF)
48-h NOEC = 5.62 mg/L (measured) (100 mg/L nominal loading rate WAF)
Supporting Source: Daphnia magna; OECD TG 202.
48-h EL50 >100 mg/L loading rate WAF
48-h NOELR = 100 mg/L
Target: No data
Toxicity to aquatic algae:
Primary source: Pseudokirchneriella subcapitata; OECD TG 201
Results based on initial nominal loading rates (WAFs):
72 -h ErL50 > 100 mg/L; 72 -h EγL50 = 37.8 mg/L; 72 -h EbL50 = 36.51 mg/L
72 -h NOErLR = 12.5 mg/L; 72 -h NOEγLR = 6.25 mg/L; 72 -h NOEbLR = 6.25 mg/L
Results based on initial measured concentrations:
72 -h ErC50 > 28.92 mg/L; 72 -h EγC50 = 15.70 mg/L; 72 -h EbC50 = 15.29 mg/L
72 -h NOErC = 6.53 mg/L; 72 -h NOEγC = 3.71 mg/L; 72 -h NOEbC = 3.71 mg/L
Supporting source:
72-h ErL50 = 49 mg/L loading rate WAF
72-h NOErLR = 25 mg/L
Target: No data
Reason / purpose:
read-across source
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence limits not stated
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95 % CL not stated

Description of key information

Oncorhynchus mykiss; OECD TG 203; read-across to EC 235-741-0.

96-h LL50 value of greater than 100 mg/L loading rate WAF

Key value for chemical safety assessment

LC50 for freshwater fish:
100 mg/L

Additional information

No study is available for Phosphoric acid, linear and branched Octyl Esters. Results are read across from an OECD TG 203 study performed on the 2-ethylhexyl- analogue. In this study, groups of 7 rainbow trout (Oncorhynchus mykiss) were exposed, in groups of seven, to a Water Accommodated Fraction (WAF) of the test item, at a single nominal loading rate of 100 mg/l for a period of 96 hours 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. The 96-h LL50 based on nominal loading rates was > 100 mg/l loading rate WAF and correspondingly the No Observed Effect Loading rate was 100 mg/l loading rate WAF.