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Short-term toxicity to fish

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Endpoint:
fish embryo acute toxicity (FET)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Initiation date: 29/01/2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with GLP.
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 236 (Fish Embryo Acute Toxicity (FET) Test)
Principles of method if other than guideline:
The test method is intended to determine the acute or lethal toxicity of the substance on embryonic stages of fish.
Newly fertilised fathead minnow eggs were exposed to the test chemical for a period of 120 hrs during a semi-static renewal test. The test includes five increasing concentrations of the chemical tested and a negative control. Every 24 hours, four apical observations are recorded as indicators of lethality: (i) coagulation of fertilised eggs, (ii) lack of somite formation, (iii) lack of detachment of the tail-bud from the yolk sac, and (iv) lack of heartbeat. At the end of the exposure period, acute toxicity is determined based on a positive outcome in any of the four apical observations recorded, and the LC50 is calculated.
OECD Test guideline 236 sets some parameters that are specific to zebrafish. This test was conducted with fathead minnows and consequently, there are some minor differences in terms of temperature and pH which are not considered to affect the reliability of the study. OECD 236 requires the use of a positive control which was not used in this test (there was no positive control carried out in this study. The statistical means and variances of performance of specific reference substances in P. promelas is less well established as yet than it is for Danio rerio. Therefore, so as to not inappropriately communicate an undue degree of precision, it was decided to rely on the control condition and development observations only, during this test.). OECD TG 236 recommends a spacing factor 1.5 - 2.2 between concentration levels; in this test a spacing factor 2.5 was used. This does not affect the reliability of the study or statistical interpretation.
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Nominal: Negative Control, Internal Control, 0.51, 1.3, 3.2, 8.0 and 20 mg/l
Mean Measured: < MQL1, < MQL1, 0.49, 1.1, 2.8, 8.5 and 20 mg/l.
- Sampling method: Water samples were collected from the newly prepared batch of test solutions of each treatment and control group at the beginning of the test on Day 0 and from old test solutions pooled from individual wells of the surrogate test chambers of each treatment and control group at 24 hours (±1 hour) to represent concentrations in the test system during each 24-hour renewal interval. Additional samples of the newly prepared batch of test solutions of each treatment and control group on Day 1 of the test were analyzed to confirm the analytical results of the new solutions collected on Day 0 of the test. Since the additional samples of new solutions collected on Day 1 of the test confirmed the measured concentrations of the new solutions on Day 0 of the test, the analytical results for the new solutions from Day 1 of the test were not used in the calculation of mean measured test concentrations. Additional samples of the old test solutions pooled from individual wells of the surrogate test chambers of each treatment and control group at 48 hours (±1 hour) were stored for possible future analysis. The samples of new solutions were collected from mid-depth, while the samples of old solutions were pooled from the test chambers. Sample collection from the 20 mg/l treatment group was discontinued after 48 hours due to 100% mortality.
- Sample storage conditions before analysis: Samples were placed in glass French squares and processed immediately for analysis or stored under refrigerated conditions prior to analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test solutions were prepared six times. The solutions at the first preparation were prepared one day prior to test initiation and were used for conditioning of glassware and test chambers. Test solution concentrations were not adjusted for the active ingredient of the test substance during preparation, and are based on the density of the test substance (0.829 g/ml). Solutions were prepared at nominal concentrations of 0.51, 1.3, 3.2, 8.0 and 20 mg/l. At each preparation, test solutions for the 8.0 and 20 mg/l test concentrations were prepared by diluting a calculated volume of test substance in dilution water (UV sterilized well water) to achieve a final volume of 1000 ml. An aliquot of the test solution at the 8.0 mg/l treatment concentration was diluted to prepare 1000 ml of the 3.2 mg/l treatment concentration. An aliquot of the test solution at the 3.2 mg/l treatment concentration was diluted to prepare 1000 ml of the 1.3 mg/L treatment concentration. Finally an aliquot of the test solution at the 1.3 mg/l treatment concentration was diluted to prepare 1000 ml of the 0.51 mg/l treatment concentration. Test solutions at the 8.0 and 20 mg/l treatment concentrations were sonicated for approximately 25 minutes and followed by inversion at least 20 times to mix. The remaining test solutions were mixed by inversion at least 20 times. The internal and negative control solutions were dilution water only. New test solutions were prepared daily. When 100% mortality occurred in the 20 mg/l treatment concentration on Day 2 of the test, the preparation of test solution at the 20 mg/l treatment concentration was discontinued.
At the beginning of the test, prior to the addition of the embryos, 2.0 mL of newly prepared test solutions and dilution water were added to the appropriate wells of each well plate using a pipette. At each renewal at approximately 24-hour intervals, old solutions of each treatment and control groups were removed from each well using a pipette. A small amount of old solution was left in each well to ensure that embryos remain covered with old test solutions to avoid drying of embryos. New solution was then added to each respective well.
One day prior to test initiation, 1000 ml of each decanol treatment concentration and control solutions were prepared and 2.0 ml of each solution was pipetted into each appropriate well and allowed to condition overnight. A portion of the appropriate test solution was also poured into Petri dishes that were conditioned for use in holding embryos prior to transfer to well plates. All remaining test solutions were held in their respective volumetric flasks to allow for conditioning glassware to be used for preparation of test solutions, prior to test initiation. Each plate was covered during the exposure period.
- Eluate:
- Differential loading:
- Controls: Dilution water.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No
Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: Fathead minnow
- Strain:
- Source: Wildlife International, Easton, Maryland 21601
- Age at study initiation (mean and range, SD): Embryo (no older than 16-celled blastodisc developmental stage)
- Length at study initiation (length definition, mean, range and SD): Not reported
- Weight at study initiation (mean and range, SD): Not reported
- Method of breeding: The fish used to supply embryos for the test were approximately 7 months of age and were sexually dimorphic adults that are reproductively mature and actively spawning. Breeding groups of two males and four females were used to supply embryos, with a total of 16 breeding groups. The fish were obtained Osage Catfisheries, Inc. of Osage Beach, Missouri. The identity of the species was verified by the supplier using appropriate taxonomic keys.
Prior to the onset of light on the day of exposure initiation, clean spawning substrates (inverted semi-circular sections of aged PVC pipe) were placed in 16 breeding tanks each containing 2 males and 4 females. After allowing sufficient time for egg deposition and fertilization, tiles containing newly deposited eggs were collected from five breeding tanks. The eggs were gently removed from the tiles and pooled. The fertilization of the embryos wasere confirmed microscopically and the fertilization rate was approximately 95%. Embryos no older than 16-celled blastodisc developmental stage were selected and placed in the transfer chambers (Petri dishes) containing appropriate test solutions.
- Feeding during test: N/A


ACCLIMATION
- Acclimation period: The breeding stock of fathead minnows was held under flow-through conditions for at least 14 days prior to collection of embryos for testing, in water from the same source and at approximately the same temperature as used during the test.
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: Daily during the holding period, the breeding stock of fathead minnows were fed a commercially-prepared diet supplied by Sera North America, Inc. of Montgomeryville, Pennsylvania and supplemented with brine shrimp nauplii (Artemia sp.) hatched from the cysts supplied by INVE Aquaculture of Salt Lake City, Utah and Brine Shrimp Direct of Ogden, Utah.
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): During this 2-week period, the breeding stock fish in the lot used for the test showed no signs of disease or stress and there was <5% mortality.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
120 h
Hardness:
100-120 mg/L as CaCO3
Test temperature:
range 24.5 - 25.7 °C
pH:
range 7.9-8.7
Dissolved oxygen:
8.2 mg/l
Nominal and measured concentrations:
Nominal: Negative Control, Internal Control, 0.51, 1.3, 3.2, 8.0 and 20 mg/l
Mean Measured: < MQL1, < MQL1, 0.49, 1.1, 2.8, 8.5 and 20 mg/l.

Details on test conditions:
TEST SYSTEM
- Test vessel: Clear Costar® polystyrene 24-well well plates (125 mm x 85mm) with approximately 2 to 5 ml filling capacity per well and each well plate was equipped with a clear polystyrene lid.
- Type (delete if not applicable): closed ( clear polystyrene lid).
- Material, size, headspace, fill volume: Clear polystyrene, 2 to 5 ml filling capacity per well, 2 ml test solution per well.
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): n/a
- Renewal rate of test solution (frequency/flow rate): Daily
- No. of organisms per vessel: 1 embryo per well, 24 wells per plate
- No. of vessels per concentration (replicates): 20 replicates per treatment and negative control groups. Dilution water was then added to the remaining 4 wells in each well plate, assigned to each treatment and control group, to serve as an internal control of each plate.
- No. of vessels per control (replicates): 20
- No. of vessels per vehicle control (replicates): n/a
- Biomass loading rate:

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water used for holding and testing was freshwater obtained from a well approximately 40 meters deep located on the Wildlife International site. The well water was passed through a sand filter to remove particles greater than approximately 25 µm, and pumped into a 37,800 l storage tank where the water was aerated with spray nozzles. Prior to use in the test system, the water was filtered to 0.45 µm to remove fine particles and was passed through an ultraviolet (UV) sterilizer. The well water is characterised as moderately-hard water.
- Total organic carbon: <2 mg/l
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:152-172 mg/l as CaCO3
- Ca/mg ratio:
- Conductivity: 245-326 (µS/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: Every 24 hours: Dissolved oxygen and pH were measured in the newly prepared test solutions immediately prior to filling the test chamber wells at the beginning of the test and at each renewal interval, and were measured in pooled solution collected from the wells in the surrogate test chambers at the end of each renewal interval and at the end of the test.
Hardness, alkalinity and specific conductance in the dilution water were measured in the negative control (dilution water) and in the highest concentration treatment group at the beginning and end of the first and last renewal periods (i.e., new solutions at 0 and 96 hours; old solutions at 24 and 120 hours).

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours of light and 8 hours of darkness. A 30 minute transition period of low light intensity was provided when lights went on and off to avoid sudden changes in light intensity.
- Light intensity: The test systems were illuminated using fluorescent tubes that emit wavelengths similar to natural sunlight.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations of mortality, abnormal development or other observed effects, and hatching were recorded for each tested embryo at approximately 24-hour intervals during the exposure period (with the exceptions indicated below). Apical observations performed on each tested embryo including coagulation of the embryo, lack of somite formation, non-detachment of the tail and lack of heartbeat.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: approximately 2.5
- Justification for using less concentrations than requested by guideline: n/a
- Range finding study
- Test concentrations: 0.25, 0.74, 2.2, 6.7 and 20 mg/l.
- Results used to determine the conditions for the definitive study: Percent mortality of embryos in the negative control, internal control and in the 0.25, 0.74, 2.2, 6.7 and 20 mg/l treatment groups at 120 hours was 0, 0, 5, 5, 15, 75 and 100%, respectively. The percent hatching in the negative control, internal control and in the 0.25, 0.74, 2.2, 6.7 and 20 mg/l treatment groups at 120 hours was 40, 54, 10, 35, 20, 0 and 0%, respectively. Based on the results of the preliminary range-finding test, nominal test concentrations of 0.51, 1.3, 3.2, 8.0 and 20 mg/l were selected for the definitive test.
Key result
Duration:
120 h
Dose descriptor:
LC50
Effect conc.:
3.4 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: (95% CL 2.8 - 8.3 mg/l)
Results with reference substance (positive control):
There was no positive control carried out in this study (as per OECD 236). The statistical means and variances of performance of specific reference substances in P. promelas is less well established as yet than it is for Danio rerio. Therefore, so as to not inappropriately communicate an undue degree of precision, it was decided to rely on the control condition and development observations only, during this test.
Reported statistics and error estimates:
The mortality data, based on the four apical observations (i.e. coagulation of the embryo, lack of somite formation, non-detachment of the tail and lack of heartbeat), were analyzed using the computer program of C. E. Stephan (3). The program was designed to calculate the LC50 value and the 95% confidence interval by probit analysis, the moving average method, and binomial probability with nonlinear interpolation (4, 5). Based on the mortality pattern in this study, nonlinear interpolation was used to calculate the 24-, 48-, 72-, 96- and 120-hour LC50 values and binominal probability was used to calculate the 95% confidence intervals. Due to the method used to calculate the 48- and 96-hour LC50 value, the slope of the concentration-response curve could not be calculated. The highest test concentration causing no mortality at test end and the lowest test concentration causing 100% mortality at test end were also reported.
Sublethal observations / clinical signs:

Table 1 Cumulative Mortality, Hatching and Observations

 

Mean Measured

Concentration1

(mg/L)

Number Observed in 24-Hour Period / Number Originally Exposed

(Observations)2

 

 

24 Hours

 

48 Hours3

 

 

CE

SF

TD

HB

Hatch

CE

SF

TD

HB

Hatch

 

Negative Control

0 / 20

0 / 20

--

--

--

 

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

 

Internal Control

0 / 24

0 / 24

--

--

--

 

0 / 24

0 / 24

0 / 24

0 / 24

0 / 24

 

 

0.49

0 / 20

0 /20

--

--

--

 

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

 

1.1

0 / 20

0 / 20

--

--

--

 

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

 

2.8

4 / 20

9 / 20

--

--

--

 

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

 

8.5

20 / 20

20 / 20

--

--

--

 

--3

--3

--3

--3

--3

 

 

20

20 / 20

20 / 20

--

--

--

 

--3

--3

--3

--3

--3

 

1  Test solution appearance: all clear and colorless at test initiation and termination.

2  Observations: CE = coagulation of the embryo; TD = absence of tail detachment; SF = absence somite formation; HB = absence of heart beat; Hatch = embryo has hatched; -- = no observation.

3  -- = No data due to 100% mortality.

Table 1 continued

 

Number Observed in 24-Hour Period / Number Originally Exposed

(Observations)2

 

72 hours

 

96 hours3

 

CE

SF

TD

HB

Hatch

 

CE

SF

TD

HB

Hatch

Negative Control

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

Internal Control

0 / 24

0 / 24

0 / 24

0 / 24

0 / 24

 

0 / 24

0 / 24

0 / 24

0 / 24

0 / 24

0.49

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

0 / 20

0 / 20

0 / 20

0 / 20

1 / 20

1.1

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

 

0 / 20

0 / 20

0 / 20

0 / 20

0 / 20

2.8

2 / 20

0 / 20

0 / 20

2 / 20

0 / 20

 

0 / 20

0 / 20

0 / 20

0 / 20

2 / 20

8.5

--3

--3

--3

--3

--3

 

--3

--3

--3

--3

--3

20

--3

--3

--3

--3

--3

 

--3

--3

--3

--3

--3

1  Test solution appearance: all clear and colorless at test initiation and termination.

2  Observations: CE = coagulation of the embryo; TD = absence of tail detachment; SF = absence somite formation; HB = absence of heart beat; Hatch = embryo has hatched.

3  -- = No data due to 100% mortality.

Table 1 continued

Mean Measured

Concentration1

(mg/L)

Number Observed in 24-Hour Period / Number Originally Exposed (Observations)2

 

Percent

Hatching

Per Treatment

120 Hours

 

CE

SF

TD

HB

Hatch

Negative Control

0 / 20

0 / 20

0 / 20

0 / 20

7 / 20

 

35

Internal Control

0 / 24

0 / 24

0 / 24

0 / 24

9 / 24

 

45

0.49

0 / 20

0 / 20

0 / 20

0 / 20

5 / 20

 

25

1.1

0 / 20

0 / 20

0 / 20

0 / 20

2 / 20

 

10

2.8

1 / 20

0 / 20

0 / 20

1 / 20

7 / 20

 

35

8.5

--3

--3

--3

--3

--3

 

0

20

--3

--3

--3

--3

--3

 

0

1  Test solution appearance: all clear and colorless at test initiation and termination.

2  Observations: CE = coagulation of the embryo; TD = absence of tail detachment; SF = absence somite formation; HB = absence of heart beat; Hatch = embryo has hatched.

3   -- = No data due to 100% mortality.

Table 2 Mortality in 24-Hour Period and LC50 Values Based on Mean Measured Test Concentrations

 Mean measured concentration (mg/l)    Number dead in 24 hr period / Cumulative number dead / Number originally exposed               Percent mortality per treatment   
 24 hours  48 hours  72 hours  96 hours  120 hours
 Negative control 0/0/20  0/0/20   0/0/20  0/0/20  0/0/20  0
 Internal control 0/0/24   0/0/24  0/0/20  0/0/20 0/0/20   0
 0.49 0/0/20   0/0/20  0/0/20  0/0/20  0/0/20  0
 1.1  0/0/20  0/0/20  0/0/20  0/0/20  0/0/20  0
 2.8 4/4/20   0/4/20  2/6/20  0/6/20  1/7/20  35
 8.5 20/20/20   20/20/20  20/20/20  20/20/20  20/20/20  100
 20 20/20/20   20/20/20  20/20/20  20/20/20  20/20/20  100
 LC50 (95% confidence interval) 4.0 (2.8 - 8.3)  4.0 (2.8 - 8.3)  3.6 (2.8 - 8.3)  3.6 (2.8 - 8.3)  3.4 (2.8 - 8.3)  

 

 

Validity criteria fulfilled:
yes
Remarks:
hatching rate in negative control 35%. Fathead minnow embryos can hatch within 3-4 days post-fertilization but hatching usually at 120 hrs. Test finished at 120 hrs, prior to hatching of remaining embryos.
Conclusions:
A 120-h LC50 of 3.4 mg/l based on mean measured concentrations has been determined for the effects of decanol on mortality of fathead minnow embryos.
Executive summary:

A 120-h LC50 of 3.4 mg/l based on mean measured concentrations has been determined for the effects of decanol on mortality of fathead minnow embryos.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to an appropriate national standard method. It was not compliant with GLP and though analytical monitoring is said to be conducted results are not reported.
Qualifier:
according to guideline
Guideline:
other: US EPA 1975
Deviations:
not specified
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 5 concentrations and control

- Sampling method: Concentrations of chemicals in water were measured daily in each tank throughout the test, although analysis results were not provided.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: dispersion (limit of water solubility was previously given by suppliers or determined analytically)
Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM

- Common name: fathead minnow

- Source: US EPA Environmental Research Laboratory-Duluth

- Age at study initiation (mean): 30 d

- Weight at study initiation (mean): 0.12 g

- Feeding during test: none


ACCLIMATION

- Acclimation period: fry grown in lab

- Acclimation conditions: same as test

- Type and amount of food: fresh hatched brine shrimp nauplii and dry flake Tetramin during their first week

- Feeding frequency: shrimp 2-3 times a day until 24h before the test

- Health during acclimation: good
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
42.2 mg/L as CaCO3
Test temperature:
25 +/- 1 degC
pH:
7.5
Dissolved oxygen:
>60%
Salinity:
n. a.
Nominal and measured concentrations:
control and 5 concentrations
Details on test conditions:
TEST SYSTEM

- Test vessel: test tanks

- Type of flow-through: proportional diluter system

- No. of organisms per vessel: 2

- No. of vessels per concentration: 2

- No. of vessels per control: 2


TEST MEDIUM / WATER PARAMETERS

- Source: Lake Superior water

- Metals: in ug/L: Al 1-26, Cd <0.1, Cu 0.3-3.2, Fe 2-83, Cb <0.5, Cr 2-20, Ni <0.5 Zn, 1.0-2.7, Mn 0.2-11.5, Mg 2.9-3.6

- Alkalinity:42.2 mg/L as CaCO3

- Culture medium different from test medium: no



OTHER TEST CONDITIONS

- Adjustment of pH: none

- Photoperiod:

- Light intensity:


EFFECT PARAMETERS MEASURED: mortality recorded at 1, 3, 6, 12, 24, 48, 72 and 96h.


TEST CONCENTRATIONS: 5 concentrations, plus the control
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
2.4 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
other: not specified
Basis for effect:
mortality (fish)
Details on results:
- Effect concentrations exceeding solubility of substance in test medium: water solubility of the test substance was determined prior to the initiation of the test.
Reported statistics and error estimates:
Median lethal concentrations were computed using the trimmed Spearman-Karber method on a PDP-11/70 computer.
Validity criteria fulfilled:
yes
Conclusions:
A 96h LC50 of 2.4 mg/L was determined for the effects of the test substance on mortality of Pimephales promelas.
Executive summary:

A 96h LC50 of 2.4 mg/L was determined for the effects of the test substance on mortality of Pimephales promelas.

Description of key information

Short-term toxicity to fish: weight-of-evidence: 120 h LC50 3.4 mg/l (measured) in accordance with test guideline OECD 236 (Pimephales promelas, embryo toxicity test) and 96-h LC50 2.4 mg/l (nominal) in accordance with test guideline OECD 203 (Pimephales promelas)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
2.4 mg/L

Additional information

Several values for short-term toxicity to fish are available in literature and consistently indicate an LC50 in the range 1 -10 mg/l in freshwater fish species. The result of highest reliability is a recent fish embryo toxicity test: A 120 h LC50 value of 3.4 mg/l (mean measured concentration) was determined for the effects of the test substance on mortality of Pimephales promelas embryos, supported by analytical monitoring and conducted in accordance with GLP (Wildlife International, 2015).

 

A 96h LC50 value of 2.4 mg/l was determined for the effects of the test substance on mortality of Pimephales promelas (Veith, et al., 1983, Brooke et al., 1984). This study reflects the lowest reliable experimental value that is available for this endpoint within the data set.

 

These results are supported by consistent literature values in several freshwater fish species and are also consistent with QSAR predictions.

 

96-h LC50 S. gairdneri (new name: O. mykiss) = >4.2 - <5.6 mg/l [EG&G Bionomics, 1975]

96-h LC50 L. macrochirus = 5.05 mg/l [EG&G Bionomics, 1975]

96-h LC50 O. mykiss = 5.7 mg/l [Huntingdon Life Sciences, 1996h]

96-h LC50 A. alburnus = 7.2 mg/l [Linden et al., 1979, Bengtsson, Renberg and Tarkpea, 1984]

48-h LC50 L. idus = 8.4 mg/l [Henkel, 1999n, rel. 4]

48-h LC50 L. idus = 0.6-3.2 mg/l [Verschueren, 1996, rel. 4]

 

96-h LC50 (fish) = 1.9 mg/l (QSAR) [Fisk et al., 2009]

 

Discussion of trends in the Category of C6-24 linear and essentially-linear aliphatic alcohols:

Linear LCAAs

The data summarised in the table below show that the toxicity of the single carbon number chain length LCAAs increases from an LC50 of 97 mg/L for C6 to 1.0 mg/L for C12. At higher carbon number chain lengths there is an absence of short-term toxicity (LC50 values are reported as being greater than the highest test concentration or higher than the water solubility of the test substance) and this is explained by the water solubility of an LCAA limiting its bioavailability, such that a toxic concentration for short-term exposure is not achieved.

The results of a 7-day, semi-static toxicity test with 1-octanol using Pimephales promelas larvae aged 1, 4 and 7 days at the start of the study have also been reported by (Pickering et al., 1996) but are not included in the table. NOECs based on nominal concentrations were 1.5-11.9 mg/L for survival. Measured concentrations in the test were reduced to non-detectable levels in most test vessels in the old media. The poor maintenance of exposure concentrations means that the reported NOEC values are almost certainly underestimates of the true toxicity of the substance. The results of this test are also discussed later with respect to long-term toxicity.

Multi-constituent LCAAs

The data for multi-constituent substances of different carbon chain length LCAAs (commercial products) are shown in Table 7.2. The results show that substances containing LCAAs with carbon numbers in the ranges of C8-10 and C6-12 exert short-term toxic effects at concentrations of between 0.7 and 10 mg/L. At these concentrations all the constituents are likely to have been fully dissolved.

In contrast, multi-constituent substances - Alcohols, C12-13; Alcohols, C12-13-branched and linear and Alcohols, C12-15-branched and linear - exhibited effects at loading rates where not all constituents were fully dissolved. Under such circumstances the presence of retained undissolved test material, such as occurred in the Shell Toxicology Laboratory (1978a) test, opens up the possibility for physical fouling of the test organism and this needs to be kept in mind when interpreting the result. The multi-constituent substances containing LCAAs with carbon chain length C12 and above did not exhibit short-term toxicity effects at loading rates where the solubility of the constituent LCAAs was exceeded.

The data for nonanol, branched and linear, decanol branched and linear, decanol branched and undecanol branched alcohols, have been read-across from their linear alcohols counterparts (C9, C10 and C11) since they are essentially linear alcohols.

Alcohols, C14-15 ecotoxicity assessment is based on weight of evidence from two studies; Bridie et al., 1973 and Shell Toxicology Lab (1978). Both studies report the LC50 to be above the limit of solubility. The Bridie et al. study tested the toxicity of the substance via the WAF preparation method (the preferred method of testing with poorly soluble mixtures) however it does not report complete information on the study methods and conditions. The Shell Toxicology Lab (1978) did not utilise WAF methods but it is reported more comprehensively.

The results for both single carbon number LCAAs and the multi-constituent substances indicate that, for fish, there is a short-term toxicity cut-off for LCAAs with carbon numbers >C14.

The lowest reliable LC50 values determined in tests with single carbon chain length LCAAs are shown in the following table. 


Table 7.1: Key fish short-term toxicity studies on single carbon chain length linear LCAAs.

CAS

Chemical Name

Comments

Water solubility (mg/L)

Species

Method/ Guideline

Exposure regime

Endpoint

Value (mg/L)1,2

Reliability code

Reference

111-27-3

1-Hexanol

 

5900 at 20°C

Pimephales promelas

US EPA 1975

Flow-through

96 h LC50

97 (m)

2

Veith, Call and Brooke, 1983a,b

111-70-6

1-Heptanol

Supporting

1300 at 20°C

Pimephales promelas

ASTM 1980

Flow-through

96 h LC50

38 (m)

2

Broderius and Kahl, 1985

111-87-5

1-Octanol

 

550 at 25°C 

Pimephales promelas

ASTM 1980

Flow-through

96 h LC50

13 (m)

2

Veith, Call and Brooke, 1983a,b; University of Wisconsin-Superior., 1984; Broderius and Kahl, 1985

143-08-8

1-Nonanol

 

130 at 20 °C

Pimephales promelas

ASTM 1980

Flow-through

96 h LC50

5.5 (m)

2

Broderius and Kahl, 1985

112-30-1

1-Decanol

 

40

Pimephales promelas (embryo)

OECD TG 236

Flow-through

120 h LC50

3.4 (m)

1

Wildlife International, 2015b

112-30-1

1-Decanol

 

40

Pimephales promelas

US EPA 1975

Flow-through

96 h LC50

2.3 (m)

2

Veith, Call and Brooke, 1983a,b; Brooke et al., 1984

112-42-5

1-Undecanol

 

8.0 at 20°C

Pimephales promelas

US EPA 1975

Flow-through

96 h LC50

1.0 (m)

2

Veith, Call and Brooke, 1983a,b

112-53-8

1-Dodecanol

 

1.9 at 20°C

Pimephales promelas

US EPA 1975

Flow-through

96 h LC50

1.0 (m)

2

Veith, Call and Brooke, 1983a,b

112-70-9

1-Tridecanol

Supporting

0.38 at 20°C

Pimephales promelas

US EPA 1975

Flow-through

96 h LC50

>0.33 (m)

2

Veith, Call and Brooke, 1983a,b

112-72-1

1-Tetradecanol

 

0.19 at 25°C

Salmo gairdneri i3

OECD 203

Semi-static

96 h LC50

>1 (n) (>LoS)

2

SafePharm, 1996b

36653-82-4

1-Hexadecanol

 

0.024 at 25°C

Salmo gairdneri3

OECD 203

Semi-static

96 h LC50

>0.4 (n) (>LoS)

2

SafePharm, 1996c

112-92-5

1-Octadecanol

 

0.0011 at 25°C

Salmo gairdneri3

OECD 203

Semi-static

96 h LC50

>0.4 (n) (>LoS)

2

SafePharm, 1996d

661-19-8

1-Docosanol

 

approx. 0.001 (estimate)

Oncorhynchus mykiss

OECD 203

Semi-static

96 h LC50

>1000 (n)
(>LoS)

2

SafePharm, 2000

Notes:

1 >LoS: concentration/Loading rate greater than the limit of water solubility

2 (n) based on nominal concentrations, (m) based on measured concentrations.

3 Now known asOncorhynchus mykiss. The names used in the study reports are given here.

 

The lowest reliable LC50 values determined in tests with multiconstituent carbon chain length LCAAs are shown in the following table.

 

Table 7.2: Fish short-term toxicity studies on mixed carbon chain length LCAAs.

CAS #

Chemical name

Comments1

Water solubility (mg/L)

Species

Method/ Guideline2

Exposure regime

Endpoint

Value (mg/L)3

Reliability code

Reference

n/a

Alcohols, C7-9

 SUPPORTING

510 at a loading rate of 1000 mg/L (estimated)

I. idus4

Not specified

Static

96 h LC50

0.7-0.8 (n)

4 (disregarded)

Shell, 1978

67762-41-8

Alcohols, C8-10

Type C

SUPPORTING

2.4 at 25°C

Salmo gairdneri4 and Lepomis macrochirus

EPA 1975

Static

96 h LC50

6.5-10 (n)

2

EG&G Bionomics, 1975

66455-17-2

Alcohols, C9-11

 SUPPORTING

44 at a loading rate of 1000 mg/L. (estimated)

S. gairdneri4

Not specified

Static

96 h LC50

6.3-10 (n)

2

Shell Toxicology Laboratory, 1979

66455-17-2

Alcohols, C9-11

 SUPPORTING

44 at a loading rate of 1000 mg/L. (estimated)

Scopthalmus maximus (marine species)

Not specified

Semi-static

96 h LC50

5.8 (n)

2

Huntingdon Life Sciences Ltd., 1991d

68515-81-1

Nonanol, branched and linear

 

121 (estimated)

Pimephales promelas

ASTM 1980

Flow-through

96 h LC50

5.5 (m)

 

(r-a from C9)

2

Broderius and Kahl, 1985

90342-32-8

Decanol, branched and linear

 

26.17 at 20°C

P. promelas

US EPA 1975

Flow-through

96 h LC50

2.3 (m)

 

(r-a from C10)

2

Veith, Call and Brooke, 1983a,b; University of Wisconsin-Superior, 1984

128973-77-3

Undecanol, branched and linear.

Reaction mass of 2-methyldecan-1-ol and 2-propyloctan-1-ol and 2-ethylnonan-1-ol and 2-butylheptan-1-ol

 

6.3 at 25°C

P. promelas

US EPA 1975

Flow-through

 96 h LC50

1.0 (m)

(r-a from C11)

2

Veith, Call and Brooke, 1983a,b

75782-86-4

Alcohols, C12-13

 

2.4 at 25oC

S. gairdneri4

Not specified

Static

96 h LC50

4.0-10 (>LoS)

2

Shell Toxicology Laboratory, 1978a

75782-86-4

Alcohols, C12-13

 

2.4 at 25oC

S. maximus (marine species)

Not specified

Semi-static

96 h LC50

10 (n) (>LoS)

2

Huntingdon Life Sciences Ltd., 1991c

740817-83-8

Alcohols, C12-13-branched and linear

 

2.9-3.1 at 20°C

Brachydanio rerio

OECD 203 WAF

Semi-static

96-hr LL50

15 (n) (>LoS)

1

TNO, 2000a

90604-40-3

Alcohols, C12-15-branched and linear

 

0.80 at 20°C

Oncorhynchus mykiss

OECD 203 WAF

Semi-Static

96 h LL50

100-300 (n) (>LoS)

1

Shell Global Solutions, 2000

68855-56-1

Alcohols, C 12-16

Type B

SUPPORTING

0.80 at 20°C

O. mykiss

Not specified

Static

96 h LC50

57 (n) (>LoS)

2

Huntingdon Life Sciences 1996i

80206-82-2

Alcohols, C 12-14

not possible to determine compositional type

SUPPORTING

approx. 4 predicted at 1000 mg/L loading rate

L. idus

OECD 203

Static

48 h LC50

>5000 (n) (>LoS)

2

Henkel, 1999m

75782-87-5

Alcohols, C14-15

 

0.7 at 20°C and 0.15 at a loading rate of 1000 mg/L. (estimated)

S. gairdneri4

Not specified

Static

96 h LL50

>500 (n) (>LoS)

2

Shell Toxicology Lab 1978b

75782-87-5

Alcohols, C14-15

 

0.7 at 20°C and 0.15 at a loading rate of 1000 mg/L. (estimated)

Carassius auratus

Not specified

Static

96 h LL50

>0.7 (n)

(>LoS)

2

Shell Internationale Chemie, 1973

68002-94-8

Alcohols, C 16-18 and 18 Unsaturated 

SUPPORTING

0.0404 predicted at 1000 mg/L loading rate

L. idus

OECD 203

Static

48 h LC50

>10000 (>LoS)

4

Henkel, 1999o

Notes:

1 Compositional Types are described in section 1.5 of the ecotoxicity category report.

2 WAF denotes test medium was a water-accommodated fraction      

3 >LoS: LC50 observed was greater than the limit of solubility of at least some constituents of the substance. (n) based on nominal concentrations, (m) based on measured concentrations.

4 Now known as Oncorhynchus mykiss. The names used in the study reports are given here.