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Long-term toxicity to fish

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Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented peer-reviewed publication.
Qualifier:
equivalent or similar to guideline
Guideline:
other: APHA 1975
GLP compliance:
no
Analytical monitoring:
not specified
Vehicle:
not specified
Test organisms (species):
other: Tilapia mossambica
Details on test organisms:
TEST ORGANISM
- Source: local farms
- Weight at study initiation (mean and range, SD): 0.786 g
- Length at study initiation (length definition, mean, range and SD): 35 mm
- Feeding during test
- Food type: 1:1 mixture of rice bran and mustard cake
- Frequency: daily


ACCLIMATION
- Acclimation period: 168 hrs
Test type:
other: static, open-system
Water media type:
freshwater
Limit test:
no
Total exposure duration:
90 d
Hardness:
hardness 290 mg/L as CaCO3
Test temperature:
temperature 27.9 ± 0.14 °C
pH:
pH 7.1 ± 0.1,
Dissolved oxygen:
dissolved oxygen 10 mg/L
Nominal and measured concentrations:
Test concentrations were 0.0, 0.25, 0.38, 0.51, and 1.10 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel: Tests were conducted in outdoor earthen vats (62 cm diameter, 30 cm mean depth)
- Renewal rate of test solution (frequency/flow rate): 15 days
- No. of organisms per vessel: 15
- Material, size, headspace, fill volume: 60-L of borehole water and 5 kg of uncontaminated soil

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Borehole water is unchlorinated water


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : feeding, growth rate, fecundity, and yield.

Reference substance (positive control):
not specified
Duration:
90 d
Dose descriptor:
NOEC
Effect conc.:
0.25 mg/L
Basis for effect:
other: Feeding, growth rate, fecundity, yield
Duration:
90 d
Dose descriptor:
LOEC
Effect conc.:
0.51 mg/L
Basis for effect:
other: Feeding, growth rate, fecundity, yield
Details on results:
The feeding rates decreased significantly at 0.25, 0.38 and 1.10 mg/L.  Fish showed erratic behaviour, irregular opercular movement, and at higher concentrations, blood exuded from the base of the pectoral and pelvic fins and head.  No apparent difference in condition factor (K) was observed at any concentration.  The maturity index (MI) of both male and female fish appeared to decrease at all concentrations, but the biological significance of this is questionable because historic control values for this parameter were not provided and the magnitude of the response did not increase with dose.  Fecundity decreased at 0.51 mg/L but not at 1.10 mg/L.  The gastrosomatic index (GSI) was significantly different at 0.51 and 1.10 mg/L. Based on the most reliable endpoints (GSI and fecundity), the NOEC would be 0.38 mg/L and the LOEC would be 0.51 mg/L.  However, the study is incompletely documented, so details of the test substance composition and testing procedure are uncertain.  True replicates were not used so statistics can not be validly conducted, though they are reported by the authors.  In view of these limitations, and previous evaluations of the study which have reported a NOEC of 0.25 mg/L (van de Plassche et al., 1999), a conservative (protective) NOEC for this study is 0.25 mg/L.
Conclusions:
The 90-day NOEC for fish was 0.25 mg/L. The 90-day LOEC for fish was 0.51 mg/L.
Executive summary:

The long-term toxicity of the test substance to fish was determined. Groups of 15 fish were exposed to concentrations of 0.0, 0.25, 0.38, 0.51, and 1.10 mg/L for 90 days. Test solutions were renewed every 15 days. The 90 -day NOEC was 0.25 mg/L, and the LOEC was 0.51 mg/L.

Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented peer-reviewed publication.
Qualifier:
according to guideline
Guideline:
other: Dutch Standard Organization NEN 6505
GLP compliance:
no
Analytical monitoring:
not specified
Test organisms (species):
Poecilia reticulata
Details on test organisms:
TEST ORGANISM
- Age at study initiation (mean and range, SD): 3 to 4 weeks
- Food type: TetraMin, TetraPhyll
Test type:
semi-static
Water media type:
freshwater
Total exposure duration:
28 d
Hardness:
measured
Test temperature:
23 ± 2 °C
pH:
measured
Dissolved oxygen:
measured
Nominal and measured concentrations:
not reported
Details on test conditions:
TEST SYSTEM
- Material, size, headspace, fill volume: 10-L per chamber
- Renewal rate of test solution (frequency/flow rate): renewed three times per week
- No. of vessels per concentration (replicates): Fifty fish were used per group

OTHER TEST CONDITIONS
- Photoperiod: circadic


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality and immobility
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Details on results:
The only effect (98% mortality at 10 mg/L) occurred within 2 days of study initiation.

The NOEC normalized by van de Plassche et al. (1999) to LABS Na was 3.2 mg/L. 

Conclusions:
The 28-day NOEC for fish was 3.2 mg/L (nominal). The 28-day LOEC was 10 mg/L (nominal).
Executive summary:

Groups of 50 Poecilia reticulata were exposed to various concentrations of the test substance for 28 days. Test solution was renewed three per week. The 28-day NOEC for fish was 3.2 mg/L (nominal). The 28-day LOEC was 10 mg/L (nominal).

Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Basic data given: comparable to guidelines/standards, with analytical. Not enough details to warrant Klimisch 1 rating.
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)
Deviations:
no
GLP compliance:
no
Remarks:
the study pre-dates GLPs
Analytical monitoring:
yes
Details on sampling:
Please see the section below. The study report does not include details about sampling.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION: Preparation of test solutions is not reported. The test concentrations were delivered to the duplicate test vessels by a modified Mount and Brungs 1L proportional diluter.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: Bluegill fish
- Strain: Not reported
- Source: Not reported
- Length at study initiation: 2.5-5.0 cm
- Age at study initiation: Juvenile
- Weight at study initiation: Not reported
- Method of breeding: Not applicable

FEEDING DURING TEST
- Food type: Frozen brine shrimp
- Amount: Fed at approx. 2% of total body weight/day
- Frequency: Daily
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Post exposure observation period:
None
Hardness:
120 mg/L CaCO3
Test temperature:
21 ± 2 °C
pH:
8.1 ± 0.2
Dissolved oxygen:
Mean Dissolved Oxygen Concentration (DOC): 7.8 mg/L (ranged, 7.0-9.0)
Nominal and measured concentrations:
Nominal test concentrations were: 0, 0.5, 1.0, 2.0, and 4.0 mg/L. MBAS and radiometric counts confirmed the expected nominal concentrations. Two figures in the study report show the confirmation of nominal test concentrations, but details not provided.
Details on test conditions:
EST SYSTEM
- Test vessel: Aquaria
- Type: Open
- Material: 19 L glass
- Aeration: No (test was flow-through)
- Type of flow-through: Proportional diluter
- Renewal rate of test solution: 8 replacement volumes/d
- No. of organisms per vessel: Five
- No. of vessels per concentration (replicates): Two
- No. of vessels per negative control (replicates): Two
- No. of vessels per vehicle control (replicates): Not applicable
- Biomass loading rate: Not reported. There were ¿5 pre-weighed juvenile bluegills per each aquarium,¿ according to the study report. Loading rate was therefore 1 juvenile bluegill/4 L test solution.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was carbon and reverse-osmosis filtered well water of 120 mg/L CaCO3 hardness.
- Total organic carbon: Not reported
- Particulate matter: Not reported
- Metals (mg/L): Zinc: <0.001; Lead: <0.01; Iron: <0.05; Copper: <0.001,
- Nitrate (mg/L): <0.05
- Pesticides: Not reported
- Chlorine: Not reported
- Alkalinity: Not reported
- Ca/mg ratio: Not reported
- Conductivity: Not reported
- Salinity: Not applicable
- Culture medium different from test medium: Not reported
- Intervals of water quality measurement: Twice weekly
- Other:
- Temperature was monitored continuously with a recording thermograph

OTHER TEST CONDITIONS
- Adjustment of pH: Not reported
- Photoperiod: Not reported
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED: (1) The growth of juvenile bluegills was determined from 10 individuals at each treatment concentration measured and weighed (wet weight) before and after the exposure. (2) Amount of 14C-LAS taken up by the fish was measured by combustion analysis.

NEGATIVE CONTROL PERFORMED: Yes (received only filtered well water)

SAMPLING
- Sampling frequency: Only on Day 28, as sampling of test species were not carried out during exposure period, to avoid disturbance of the developing fishes. On Day 28, all 10 fish per test concentration were recovered.
- Sampling method: For the amount of 14C-LAS taken up by the fish, triplicate samples of the test species were taken at 28 d sampling period, rinsed, dried at 30°C for 24 h, and weighed in tared plancets prior to combustion analyses for radiometric counting of 14C-LAS content
- Sample storage conditions before analysis: For growth rate, fish were weighed immediately after conclusion of test. For 14C-LAS analysis, samples were dried at 30°C for 24 h before analysis

RANGE-FINDING STUDY: No
Reference substance (positive control):
no
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
growth rate
Remarks on result:
other: Nominal concentration confirmed by analytical measurement
Details on results:
- Fish weights (individual and mean values) on day 0, 14 (if measured) and 28: Not reported
- Tank-average or pseudo-specific growth rates for periods 0-28 or 0-14 and 0-28 (if possible): Not reported
- Type of and number with morphological abnormalities: Not reported
- Type of and number with behavioural abnormalities: Not reported
- Other biological observations: Fish growth was reduced at both 2.0 and 4.0 mg/L test concentrations with no effect at 1.0 mg/L. At the end of the exposure period, test species had a biomass of 44 and 10.5 g/m2 at test concentration of 1 and 2 mg/L respectively. Control fish production was 42.5 g/m2
- Bioconcentration Data: The BCF value was found to be 252 based on a wet weight basis
- Effect concentrations exceeding solubility of substance in test medium: No
- Incidents in the course of the test which might have influenced the results: Not reported
Results with reference substance (positive control):
Positive control was not included in the study
Reported statistics and error estimates:
Not reported
Validity criteria fulfilled:
yes
Conclusions:
The 28 d NOEC of C12 LABS Na to juvenile Bluegills (Lepomis macrochirus) was 1 mg/L, based on growth rate in a flow-through model ecosystem study. Analytical measurements confirmed the nominal test concentrations.
Executive summary:

A long term toxicity test to fish (juvenile Lepomis macrochirus) was conducted on C12 LABS Na (linear alkylbenzene sulfonate),following the OECD 204 guideline. The growth of juvenile bluegills was determined after 28 days in a flow-through model ecosystem study.  The nominal test concentrations were 0, 0.5, 1.0, 2.0, and 4.0 mg/L, confirmed by analytical measurements.

The growth of juvenile bluegills (wet weight after 28 d exposure) was not affected at 0.5 and 1.0 mg LABS Na/L, but was reduced at 2.0 and 4.0 mg/L.  At the end of the exposure period, fish at 1.0 mg/L LABS Na had a biomass of 44 gm/m2 compared to 10.5 gm/m2 for the 2.0 mg/L concentration, and 42.5 gm/m2 for the controls.

The No observed effect concentration (NOEC) was 1.0 mg/L, based on growth rate.

This Chronic toxicity test is classified as acceptable, and satisfies the guideline requirements for the OECD 204.

Endpoint:
fish short-term toxicity test on embryo and sac-fry stages
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Nov. 27, 1986-Feb. 6, 1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
no guideline available
Principles of method if other than guideline:
Fertilized eggs of Salmo gairdneri were exposed to concentrations of 0.03, 0.23, 0.35, 0.63, 0.95 and 1.9 mg/L (mean measured), for 70 days. The responses recorded included the survival of eggs, time to eyed egg stage, time to hatch, survival and final weight of sac-fry (eleutheroembryos), time and extent of swim-up (external feeding)
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Representative test vessels were sampled six times during the study, and two days before introduction of eggs.
Details on test solutions:
A stock solution of 920 mg LABS Na/L was prepared every seven days. Test solutions were made by a mini-diluter system.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Eggs and sperm from Chirk trout hatchery, North Wales, were obtained from 6 individual, over 4 year old, rainbow trout and transported to the laboratory where the eggs were fertilized less than 4 hours after stripping. After swim-up, each fish was fed commercial trout feed at 8% body weight per day spread over 2-3 feedings.
Test type:
flow-through
Water media type:
freshwater
Total exposure duration:
72 d
Post exposure observation period:
Exposure was continued until several days post-swim-up, 72 days post-fertilization. The responses recorded included the survival of eggs, time to eyed egg stage, time to hatch, survival and final weight of sac-fry (eleutheroembryos), time and extent of swim-up (external feeding).
Hardness:
70-133 mg/L CaCO3
Test temperature:
8.5-11.5 °C
pH:
7.3-7.9
Dissolved oxygen:
8.8-10.3
Nominal and measured concentrations:
Nominally: 0, 0.2, 0.35, 0.62, 1.12 and 2.0 mg/L.
Mean measured concentrations: 0.03, 0.23, 0.35, 0.63, 0.95 and 1.9 mg/L.
Details on test conditions:
Fertilized eggs were exposed in a mini-diluter test system on stainless steel mesh beds in 0.5 L test vessels in two treatment groups. One group was exposured to LAS immediately post-fertilization and one group was exposured to LAS from the eyed-egg stage (19 days post-fertilization). This group was maintained in the test system and dosed with dilution water (carbon filtered tap water) prior to exposure to LAS. Each test vessel contained 30 eggs at the start of the test. Any dead life stages were removed daily.

LAS test solutions were dosed continuously at 25 mL/min, equivalent to approximately 15 volume replacements per day. There were 2 replicates for each treatment and 6 exposure concentrations.
Duration:
72 d
Dose descriptor:
NOEC
Effect conc.:
0.23 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: overall survival (fertilization to swim-up)
Details on results:
NOECs based on mean measured concentrations (mg/L) were:

time to eyed egg stage no effect at highest concentration
survival of eggs exposed from eyed stage 0.23
survival of eggs exposed from fertilization 0.23
time to hatch no effect at highest concentration
survival of sac-fry 0.23
overall survival (fertilization to swim-up) 0.23
final weight of sac-fry 0.35
time to swim-up eggs exposed from fertilization 0.35
time to swim-up exposed from eyed stage no effect at concentrations where fry survive
Conclusions:
The lowest NOEC for Salmo gairdneri over 72 days was 0.23 mg/L.
Executive summary:

Fertilized eggs of Salmo gairdneri were exposed to concentrations of 0.03, 0.23, 0.35, 0.63, 0.95 and 1.9 mg/L (mean measured), for 72 days. The responses recorded included the survival of eggs, time to eyed egg stage, time to hatch, survival and final weight of sac-fry (eleutheroembryos), time and extent of swim-up (external feeding). The lowest NOEC value found was 0.23 mg/L based on survival of eggs exposed from eyed stage, survival of eggs exposed from fertilization, survival of sac fry, and overall survival from fertilization to swim-up. The data are for C11.6 LABS Na and no normalization is required.

Endpoint:
fish life cycle toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented peer-reviewed publication.
Principles of method if other than guideline:
During the first four months a serial-dilution unit was used to maintain the LAS at the desired concentration in System I and a proportional dilution unit was used in System II, both to provide continuous exposures to fathead minnows. During the latter part of the test, the flows from a new proportional dilution unit were divided to supply both exposure systems and a fry exposure system.
GLP compliance:
not specified
Analytical monitoring:
yes
Vehicle:
not specified
Test organisms (species):
Pimephales promelas
Details on test organisms:
Fish were obtained from the ponds at the Newtown Fish Farm, Ohio Division of Wildlife. The small young-of-the-year fish were collected in late September and held in cold water. During the last week of December, 25 fish were placed randomly into each exposure chamber where they were maintained in a continuous flow of dilution water similar to that used during the test. The four largest and eight smallest fish from each aquarium were selected in order to reduce the number of fish and to try to get a more equal distribution of sexes. The exposure to LAS was started during the last week of February.
Test type:
other: other, open-system
Water media type:
freshwater
Limit test:
no
Total exposure duration:
196 d
Hardness:
water hardness ranged from 194-214 mg/L CaCO3
pH:
pH ranged from 7.50-7.95
Dissolved oxygen:
Mean dissolved oxygen 5.84-6.42 mg/L
Nominal and measured concentrations:
Test concentrations were 0.34, 0.63, 1.2 and 2.7 mg/L.
Reference substance (positive control):
not specified
Duration:
196 d
Dose descriptor:
NOEC
Effect conc.:
0.63 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
other: fry survival
Duration:
196 d
Dose descriptor:
LOEC
Effect conc.:
1.2 mg/L
Nominal / measured:
meas. (geom. mean)
Basis for effect:
other: fry survival
Details on results:
Hatchability and growth were not significantly affected. This study was on C12-LAS and when normalized to C11.6, the value is 0.90 mg/L (van de Plassche et al., 1999).
Conclusions:
The 196 day NOEC level was 0.63 mg/L. The LOEC was 1.2 mg/L.
Executive summary:

Fathead minnows were exposed to LABS Na in continuous flow systems for a total of 196 days. Results indicate that lethality of LABS Na to newly hatched fry was the most critical factor found.

Description of key information

No study was available on chronic toxicity of the test substance to fish. Therefore, read across is performed using five key studies from the related substance benzenesulfonic acid (LABS Na). The study by Pickering and Thatcher (1970) obtained a 196 day NOEC level of 0.63 mg/L. Fathead minnows were exposed to benzenesulfonic acid (LABS Na) in continuous flow systems for a total of 196 days. Chattopahyay and Konar (1985) exposed groups of 15 Tilapia mossambica to benzenesulfonic acid (LABS Na) for 90 days. The 90 day NOEC was 0.25 mg/L. The study by Canton and Sloof (1982) exposed 50 Poecilia reticulata to various concentrations of benzenesulfonic acid (LABS Na) for 28 days. The 28 day NOEC for was 3.2 mg/L (nominal). In the study of Marshall et al. (2010) fertilized eggs of Salmo gairdneri were exposed to benzenesulfonic acid (LABS Na) for 72 days and following endpoints were recorded: the survival of eggs, time to eyed egg stage, time to hatch, survival and final weight of sac-fry (eleutheroembryos), time and extent of swim-up (external feeding). The lowest 72 day NOEC value found was 0.23 mg/L. Maki et al. (1981) studied long term toxicity of benzenesulfonic acid (LABS Na) on juvenile bluegills (Lepomis macrochirus). The growth of juvenile bluegills was determined after 28 days in a flow-through model ecosystem study.  The nominal test concentrations were 0, 0.5, 1.0, 2.0, and 4.0 mg/L, confirmed by analytical measurements. The No observed effect concentration (NOEC) was 1.0 mg/L, based on growth rate.  A worst case approach was applied for chemical safety assessment and the most conservative NOEC was used (72 day NOEC of 0.23 mg/L).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
0.23 mg/L

Additional information