Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: A well documented study, but the exposure duration is shorter than the guideline requires
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
exposure duration is 24h and not 48h
GLP compliance:
not specified
Analytical monitoring:
no
Details on sampling:
- Concentrations: 5000, 2000, 1000, 100, 10 mg/L
- Sampling method: confirmation of stock solution but not of test concentrations
- Sample storage conditions before analysis: no data
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: potassium dichromate
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnia
- Strain: Straus
- Source: in laboratory breed
- Age at study initiation: >3h, <24h
- Method of breeding: adult animals were cultivated in 4L glass vessels at 21C. The medium was changed once per week. The medium was synthetic water according to DIN 38 412 Part 11. Fed twice daily with algae (Scenedesmus supspicatus).
- Feeding during test: no

ACCLIMATION
- Acclimation period: in house culture and culture medium and test dilution medium are the same
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
24 h
Hardness:
synthetic water according to DIN 38 412 Part 11
Test temperature:
21C
pH:
synthetic water according to DIN 38 412 Part 11
Dissolved oxygen:
not reported
Nominal and measured concentrations:
Nominal concentrations - 1000, 100, 10 mg/L and control in a study conducted on March 7, 1983; and 5000 and 2000 mg/L and a control in a study conducted on March 22, 1983.
No measured concentrations
Details on test conditions:
TEST SYSTEM
- Test vessel: glass
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 100 mL
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per positive control (replicates): 2
- Biomass loading rate: no data

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: synthetic water according to DIN 38 412 Part 11
- Culture medium different from test medium: no, they are the same
- Intervals of water quality measurement: no data

OTHER TEST CONDITIONS
- Light intensity: dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility at 24 hours

Reference substance (positive control):
yes
Remarks:
1.5 mg/L potassium dichromate
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
>= 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
10% immobilization in blank control in 3 concentration test conducted on March 7, 1983
15% immobilization at 10 mg/L
5% immobilization at 100 mg/L
35% immobilization at 1000 mg/L

0% immobilization in blank control in 2 concentration test conducted on March 22, 1983
0% immobilization at 2000 mg/L
10% immobilization at 5000 mg/L
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Mortality: 100% at 1.5 mg/L in both tests conducted on March 7 and on March 22, 1983
- EC50/LC50: expected to be in the range of 0.6 and 2.1 mg/L; suggesting that the daphnids used in this study were at least as sensitive and perhaps more sensitive than anticipated for this species.
Validity criteria fulfilled:
yes
Remarks:
except exposure duration less than the required 48h
Conclusions:
EC 50 48 h > 1000 mg/L corresponding to > 650 mg/L active substance.
Executive summary:

The short term toxicity to invertebrates of Toluene 4-sulphonic acid was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test. The total test period was 24 hours, a deviation respect the 48 hours reported in the guideline. A test was performed exposing daphnids to four concentration of Toluene 4-benzene sulphonic acid: 5000, 2000, 1000, 100 and 10 mg/L. The tested item did not induce acute immobilisation of Daphnia Magna at any concentration after 24 hours of exposure. Based on the reduced exposure time, this well-documented, but short-duration study provides weight of evidence that toluene-4-sulphonic acid has a relatively low toxicity to daphnids, with an EC50 > 1000 mg/L.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from a member of the Hydrotropes category
Adequacy of study:
supporting study
Study period:
March 24-26, 1993
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
no
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnid
- Source: SLI laboratory culture
- Age at study initiation (mean and range, SD): <24 hours
- Weight at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD):
- Valve height at study initiation, for shell deposition study (mean and range, SD):
- Peripheral shell growth removed prior to test initiation:
- Method of breeding: Daphnids were not fed during the test

ACCLIMATION
- Acclimation period: yes
- Acclimation conditions (same as test or not): yes
- Type and amount of food: combination of 2 mL Ankistrodesmus falcatus and 0.5 mL of trout food suspension
- Feeding frequency:
- Health during acclimation (any mortality observed): healthy
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
160 mg/L
Test temperature:
19-20 degrees C
pH:
8.2
Dissolved oxygen:
87-98 % of saturation
Nominal and measured concentrations:
0, 130, 220, 360, 600 and 1000 mg/L as active ingredient (nominal)
Details on test conditions:
TEST SYSTEM
- Test vessel Type open
- Material, size, headspace, fill volume: 250 mL glass beakers filled with 200 mL test solution
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: fortified well water following published ASTM methods and filtered through a Amberlite XAD-7 resin column to remove any organic contaminants
- Total organic carbon: 1.5 mg/L
- Metals: all less than limit of quantitation except 2.3 mg/L magnesium and 0.02 mg/L manganese
- Pesticides: all less than limit of quantitation
- Alkalinity: 90-110 mg/L as CaC03- Ca/mg ratio: 3:1
- Conductivity: 500-800 micromhos per centimeter

OTHER TEST CONDITIONS
- Photoperiod: 16 hrs light and 8 hrs dark
- Light intensity: 30-100 footcandles at the surface

EFFECT PARAMETERS MEASURED : immobilization at 24 hour intervals

TEST CONCENTRATIONS
- Spacing factor for test concentrations: <2X
- Range finding study:yes
- Test concentrations: 0.10 - 100 mg/L as active ingredient.
Following 48 hours of exposure, no mortality or sublethal effects (e.g. lethargy, erratic behavior) were onberved among any of the daphnids exposed to any concentration tested.
- Results used to determine the conditions for the definitive study: no immobilization at any test concentration
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Remarks on result:
other: no immobility observed at any test concentration
Results with reference substance (positive control):
no reference substance
Validity criteria fulfilled:
yes
Conclusions:
EC50 > 1000 mg/L active ingredient
Executive summary:



The short term toxicity of Sodium (Xylene and 4-ethylbenzene) sulfonates to Daphnia magna was assessed following EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids). Water quality parameters of temperature, dissolved oxygen, and pH were measured throughout the test and were within acceptable limits. The daphnids used for this study were « 24 hours old) and were cultured at the lab. Culture records indicated the adult daphnids were in good condition. The study was conducted with concentrations of 0, 130, 220, 360, 600 and 1000 mg/L as active ingredient (nominal). Five daphnids were added to each replicate beaker for a total of 20 daphnids per test concentration and control. The 24- and 48-hour ECso values were both > 1000 mg/L. The results indicated the 48-hour no-observed effect concentration (NOEC) was > 1000 mg/L, based on the lack of immobility or significant abnormal effects at this concentration.




Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from a member of the Hydrotropes category
Adequacy of study:
supporting study
Study period:
October 5-7, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study without complete details
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
Daphnia magna
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
270 mg/L as CaC03
Test temperature:
21 degrees Celcius
pH:
7.8 +/- 0.2
Dissolved oxygen:
approximately air saturation
Nominal and measured concentrations:
100 mg/L as test material corresponding to 40.3 mg/L active ingredient (nominal)
Details on test conditions:
TEST SYSTEM
- Test vesse Type: open
- Material, size, headspace, fill volume: 250 mL container with 200 mL test solution
- Aeration: none- Type of flow-through (e.g. peristaltic or proportional diluter):
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilization/mortality

TEST CONCENTRATIONS-
Spacing factor for test concentrations: 10X
- Range finding study: yes
Test concentrations: DRF 0.10, 1.0, 10, 100 mg/L
Results used to determine the conditions for the definitive study: no immobilization/mortality at 100 mg/L
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 40.3 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: immobilization
Details on results:
No immobilization observed in controls or at 100 mg/L test substance (or 40.3 mg/L active ingredient) The study was repeated twice (including March and June 1995 - studies #071/363 and 071/363R, respectively) with the same protocol and with the same outcome.
Validity criteria fulfilled:
not specified
Conclusions:
EC 50 > 40.3 mg/L based on active ingredient (highest dose tested)
Executive summary:

The short term toxicity to invertebrates of Sodium (Xylene and 4-ethylbenzene) sulfonates was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test. In the static limit test on Sodium (Xylene and 4-ethylbenzene) sulfonates no mortality or immobility was observed and the 48-hr EC50 was reported as > 100 mg/L (nominal concentration) which correspond to 40.3 mg/l (a.i.).

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from a member of the Hydrotropes category
Adequacy of study:
supporting study
Study period:
2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Out of the stock solution 6 concentrations were tested in a
test item solutions geometric series with a dilution factor of 2: 7.50 – 15.0 – 30.0 – 60.0 – 120 – 240 mg/L, corresponding to the nominal concentrations of 6.80 – 13.6 – 27.2 – 54.4 – 109 – 217 mg a.s./L.
The dilution levels are based on the results of a semi-static preliminary range finding test (non-GLP).

- Sampling method: At the start, samples were taken after preparation of each concentration level and the control and analyzed. At the end of the exposure (48 hours), samples of the old media were taken preferably from the test vessels

- Sample storage conditions before analysis: 18 – 25 °C, dark, in tightly closed original container
Vehicle:
yes
Details on test solutions:
In glass vessels (2 – 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, 16 hours illumination, light intensity of max. 1500 lx.

Stock solution, media 240 mg test item/L were freshly prepared with dilution water preparation. The dispersion was agitated until the solution was visually clear.

Out of the stock solution 6 concentrations were tested in a
test item solutions geometric series with a dilution factor of 2: 7.50 – 15.0 – 30.0 – 60.0 – 120 – 240 mg/L, corresponding to the nominal concentrations of 6.80 – 13.6 – 27.2 – 54.4 – 109 – 217 mg a.s./L.
The dilution levels are based on the results of a semi-static preliminary range finding test (non-GLP).
Test organisms (species):
Daphnia magna
Details on test organisms:
Test system Daphnia magna STRAUS, obtained from continuous laboratory cultures.
Reason for the selection Daphnia magna is the preferred species in accordance with the of the test system test guideline and is bred at the test facility.
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
Breeder: Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
Feeding of the culture stocks: The daphnids are fed at least 5 times per week ad libitum with a mix
of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus. The algae are cultured at the test facility.
Number of daphnids and replicates: 20 daphnids, divided into 4 replicates, each with 5 daphnids, were used for each concentration and the control.
Age of the daphnids at the start of the exposure: Less than 24 hours old daphnids from a healthy stock were used for the study. To achieve this, juvenile daphnids were removed from the
culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period were used for the test. No first brood progeny was used for the test.
Acclimatization Acclimatization of the daphnids was not necessary, because the composition of the dilution water was equivalent to the culture medium.
Application 20 g test solution per replicate were weighed out into each test vessel. This corresponds to 20 mL per test vessel. The daphnids were inserted with a small amount of dilution water by a pipette.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
see tables attached
Test temperature:
18 - 22 °C, constant within ± 1 °C
pH:
see tables attached
Dissolved oxygen:
see tables attached
Conductivity:
see tables attached
Nominal and measured concentrations:
test tiem 7.50 – 15.0 – 30.3 – 60.0 – 120 – 240 mg/L, corresponding to the nominal concentrations of 6.80 – 13.6 – 27.2 – 54.4 – 109 – 217 mg a.i./L.
Details on test conditions:
Test vessels Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses
Test volume 20 mL
Dilution water Same composition as the culture medium
Test temperature (target) 18 - 22 °C, constant within ± 1 °C
Illumination (target) Diffuse light, light intensity of max. 1500 lx
Photoperiod (target) 16/8 hours light/dark cycle

Biological Parameters
Immobilization and other observations: Immobilization was determined in all groups after 24 and 48 hours. An animal was considered immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel.
Observation of other adverse, sub-lethal effects (e.g. discoloration, abnormal behavior etc.) was done, but none occurred during the course of the study.

Water Quality Parameters
Dilution water: Prior to the start of the exposure (0 hours), the water quality parameters (i.e. pH-value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured.
Test media: At the start of the exposure (0 hours), the water quality parameters of the fresh media (i.e. pH-value, dissolved oxygen concentration) were measured in one additional replicate (without daphnids) per concentration level and control.
At the end of the exposure (48 hours), the water quality parameters of the old media were measured in one appropriate replicate (containing daphnids) per concentration level and control. The replicate with the highest immobilization rate per concentration level and control was measured.

Temperature The incubator temperature was recorded throughout the period of the test.
Quality criteria for the water quality parameters (target)
• The dissolved oxygen concentration in old media should be ≥ 3 mg/L in all concentration levels and in the control.
• The pH should be in the range of 6 – 9.
• The deviation of the final pH-values (old media) from the initial values (fresh media) should not exceed 1.5 units

Dilution water: Prior to the start of the exposure (0 hours), the water quality parameters (i.e. pH-value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured.
Test media At the start of the exposure (0 hours), the water quality parameters of the fresh media (i.e. pH-value, dissolved oxygen concentration) were measured in one additional replicate (without daphnids) per concentration level and control.
At the end of the exposure (48 hours), the water quality parameters of the old media were measured in one appropriate replicate (containing daphnids) per concentration level and control. The replicate with the highest immobilization rate per concentration level and control was measured.

Equipment
Balances SARTORIUS, METTLER and KERN
Conductometer Cond 340i (WTW)
Incubator with Timer Rumed (RUBARTH APPARATE)
Oximeter and pH-Meter HQ 40d multi (HACH LANGE)
Piston-stroke pipettes Finnpipette digital and Finnpipette F2 (THERMO SCIENTIFIC)
Spectrophotometer NANOCOLOR® UV/VIS (MACHEREY-NAGEL)
Thermohygrograph THIES
Standard laboratory equipment
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 217 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
The measured concentrations of the test item Sodium toluene-4-sulphonate in the fresh media were in the range of 99 to 107% of the nominal values. At the end of exposure after 48 hours, the measured concentrations were in the range of 96 to 106% of the nominal test item concentrations.
Results with reference substance (positive control):
The percentage of immobility for the reference item potassium dichromate (SIGMA-ALDRICH, batch number BCCC1619, purity 100.0%, CAS RN 7778-50-9) was determined after 24 hours.
The EC50 is 1.57 mg/L (validity range 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000)).

Table 1: EC10-, EC50- and EC100-Values 


 (based on the nominal concentrations of the test item and the nominal concentration of the a.s.)























Effect values



Test


duration


[hours]



Nominal concentrations of the


test item


[mg/L]



Nominal concentrations of the


active substance


[mg a.s./L]



EC10 / 50 / 100



24



> 240



> 217



48



> 240



> 217



Table 3:    Immobilization Rates after 24 and 48 hours of Exposure


              (n = 20, divided into 4 replicates with 5 daphnids each)

































































































































Nominal concentration of the test item


[mg/L]



Nominal concentration of the active substance


[mg a.s./L]



IMMOBILIZATION [%]



24 hours



48 hours



Replicates



Replicates



1



2



3



4



MV



1



2



3



4



MV



240



217



0



0



0



0



0



0



0



0



0



0



120



109



0



0



0



0



0



0



0



0



0



0



    60



    54.4



0



0



0



0



0



0



0



0



0



0



    30



    27.2



0



0



0



0



0



0



0



0



0



0



    15



    13.6



0



0



0



0



0



0



0



0



0



0



      7.5



      6.8



0



0



0



0



0



0



0



0



0



0



Control



Control



0



0



0



0



0



0



0



0



0



0



 


Table 4:  Absolute Numbers of immobile Daphnids after 24 and 48 hours of Exposure


              (n = 20, divided into 4 replicates with 5 daphnids each)

































































































































Nominal concentration of the test item


[mg/L]



Nominal concentration of the active substance


[mg a.s./L]



Number of immobile Daphnids / Total number of Daphnids



24 hours



48 hours



Replicates



Replicates



1



2



3



4



Sum



1



2



3



4



Sum



240



217



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



120



109



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



    60.0



    54.4



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



    30.0



    27.2



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



    15.0



    13.6



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



      7.50



      6.8



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



Control



Control



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



Table 5:     Measured Concentrations of the Test Item Sodium toluene-4-sulphonate during the Definitive Test




































































Sampling date



Fresh media,
0 hours



Old media,
48 hours



Nominal Concentration of the Test item


[mg/L]



Sodium toluene-4-sulphonate



Meas. conc.


[mg/L]



[%]



Meas. conc.


[mg/L]



[%]



240



256



107



251



105



120



126



105



127



106



60.0



61.6



103



61.0



102



30.0



29.7



99



28.8



96



15.0



15.2



101



15.4



103



7.5



7.90



105



7.79



104



Control



< LOQ



< LOQ



Meas. conc. = measured concentration of the test item, mean value of two replicates, dilution factors taken into account


%                   = percentage of the nominal concentration of the test item


LOQ              = limit of quantification (0.5 mg test item/L)





















Validity criteria (target)



Result



Valid



10% of the daphnids in the control were immobile or showed signs of disease or stress, e.g. discoloration or unusual behavior such as trapping on the surface of the water, during the 48-hour test period



No daphnid



yes



 3 mg O2 /L in the 48-hours old media at the end of the exposure in all concentration levels and in the control



≥ 7.55 mg/L



yes



 

Validity criteria fulfilled:
yes
Conclusions:
EC50 > 240 mg/L 48 h corresponding to > 217 mg/l active ingredient
Executive summary:

The short term toxicity to invertebrates of Sodium toluene 4-sulphonate was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test.


The study was conducted under static conditions over a period of 48 hours with six nominal concentrations of the test item  prepared in a geometric series with a separation factor of 2 (nominal concentrations of 7.50 to 240 mg/L), corresponding to 6.80 – 217 mg active ingredient/L.


All test item concentrations and the control were colorless and visually clear throughout the exposure period.


Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control.


The concentrations of the test item sodium p-cumenesulphonate were analytically verified via LCMS/ MS in the fresh media at the start of the exposure (0 hours) and in the old media at the end of the exposure (48 hours) in all concentration levels and in the control.


The measured concentrations of the test item at the start of the exposure (0 hours) were in the range of 95 to 117% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 96 to 101% of the nominal values.


The measured test item concentrations remained within ± 20% of the nominal concentrations over the test period. This indicates that the test item concentrations were stable during the duration of the test. The effect concentrations are based on the analytically verified nominal concentrations of the test item sodium toluene 4-sulphonate and the active substance concentrations.


The validity criteria of the test guideline were fulfilled.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from a member of the Hydrotropes category
Adequacy of study:
supporting study
Study period:
March 28-30, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study without complete details
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
Daphnia magna
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
270 mg/L as CaC03
Test temperature:
21 degrees Celcius
pH:
7.8+/-0.2
Dissolved oxygen:
approximately air saturation
Nominal and measured concentrations:
100 mg/L as test substance corresponding to 40.3 mg/L active ingredient (nominal)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type : open
- Material, size, headspace, fill volume: 250 mL container with 200 mL test solution
- Aeration: none
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates):-

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilization/mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10X
- Justification for using less concentrations than requested by guideline:
- Range finding study: YES with test concentrations: 0.10, 1.0, 10, 100 mg/L
- Results used to determine the conditions for the definitive study:no immobilization/mortality at 100 mg/L
Reference substance (positive control):
no
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
>= 40.3 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
other: immobilization
Validity criteria fulfilled:
not specified
Conclusions:
EC50 > 100 mg/L corresponding to 40.3 mg/L active ingredient
Executive summary:

The short term toxicity to invertebrates of Sodium p-cumenesulphonate was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test. In the static limit test on sodium cumenesulphonate no mortality or immobility was observed and the 48-hr EC50 was reported as >100 mg/L (nominal concentration) which correspond to 40.3 mg/l (a.i.).

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from a member of the Hydrotropes category
Adequacy of study:
supporting study
Study period:
2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Out of the stock solution 5 concentrations were tested in a test item solutions geometric series with a dilution factor of 2:
19.0 - 38.0 - 76.0 – 152 – 304 mg/L corresponding to the active substance concentrations:
7.54 - 15.1-30.2-60.3-121 mg a.s./L
- Sampling method: At the start, samples were taken after preparation of each concentration level and the control and analyzed. At the end of the exposure (48 hours), samples of the old media were taken preferably from the test vessels
- Sample storage conditions before analysis:18 – 25 °C, dark, in tightly closed original container
Vehicle:
yes
Details on test solutions:
Culture In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture
medium, at 20  2 °C, 16 hours illumination; light intensity of max.
1500 lx
Culture medium: Elendt M4, according to OECD 202.

Stock solution, media 400 mg/L were freshly prepared with dilution water preparation. The dispersion was agitated until the solution was visually clear.
Preparation of the test item solutions: Out of the stock solution 5 concentrations were tested in a
geometric series with a dilution factor of 2:
19.0 - 38.0 - 76.0 – 152 – 304 mg/L corresponding to the active substance concentrations:
7.54 - 15.1-30.2-60.3-121 mg a.s./L
The dilution levels are based on the results of a preliminary range finding test (non-GLP)
Test organisms (species):
Daphnia magna
Details on test organisms:
Test system Daphnia magna STRAUS, obtained from continuous laboratory cultures.
Reason for the selection Daphnia magna is the preferred species in accordance with the
of the test system test guideline and is bred at the test facility.
Origin Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
Breeder Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
Feeding of the culture stocks The daphnids are fed at least 5 times per week ad libitum with a mix
of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus. The algae are cultured at the test facility.
Number of daphnids and replicates: 20 daphnids, divided into 4 replicates, each with 5 daphnids, were used for each concentration and the control.
Age of the daphnids at the start of the exposure: Less than 24 hours old daphnids from a healthy stock were used for the study. To achieve this, juvenile daphnids were removed from the
culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period were used for the test. No first brood progeny was used for the test.
Acclimatization Acclimatization of the daphnids was not necessary, because the composition of the dilution water was equivalent to the culture medium.
Application 20 g test solution per replicate were weighed out into each test vessel. This corresponds to 20 mL per test vessel. The daphnids were inserted with a small amount of dilution water by a pipette.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
see tables attached
Test temperature:
18 - 22 °C, constant within ± 1 °C
pH:
see tables attached
Dissolved oxygen:
see tables attached
Conductivity:
see tables attached
Nominal and measured concentrations:
19.0 - 38.0 - 76.0 – 152 – 304 mg/L corresponding to the active substance concentrations:
7.54 - 15.1-30.2-60.3-121 mg a.s./L
Details on test conditions:
Test vessels Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with
watch glasses
Test volume 20 mL
Dilution water Same composition as the culture medium
Test temperature (target) 18 - 22 °C, constant within ± 1 °C
Illumination (target) Diffuse light, light intensity of max. 1500 lx
Photoperiod (target) 16/8 hours light/dark cycle

Biological Parameters
Immobilization and other observations: Immobilization was determined in all groups after 24 and 48 hours. An animal was considered immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel.
Observation of other adverse, sub-lethal effects (e.g. discoloration, abnormal behavior etc.) was done, but none occurred during the course of the study.

Water Quality Parameters
Dilution water Prior to the start of the exposure (0 hours), the water quality parameters (i.e. pH-value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured.
Test media At the start of the exposure (0 hours), the water quality parameters of the fresh media (i.e. pH-value, dissolved oxygen concentration) were measured in one additional replicate (without daphnids) per concentration level and control.
At the end of the exposure (48 hours), the water quality parameters of the old media were measured in one appropriate replicate (containing daphnids) per concentration level and control. The replicate with the highest immobilization rate per concentration level and control was
measured.

Temperature The incubator temperature was recorded throughout the period of the
test.
Quality criteria for the water quality parameters (target)
• The dissolved oxygen concentration in old media should be
≥ 3 mg/L in all concentration levels and in the control.
• The pH should be in the range of 6 – 9.
• The deviation of the final pH-values (old media) from the initial values (fresh media) should not exceed 1.5 units


Dilution water Prior to the start of the exposure (0 hours), the water quality parameters (i.e. pH-value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured.
Test media At the start of the exposure (0 hours), the water quality parameters of the fresh media (i.e. pH-value, dissolved oxygen concentration) were measured in one additional replicate (without daphnids) per concentration level and control.
At the end of the exposure (48 hours), the water quality parameters of the old media were measured in one appropriate replicate (containing daphnids) per concentration level and control. The replicate with the highest immobilization rate per concentration level and control was
measured.

Equipment
Balances SARTORIUS, METTLER and KERN
Conductometer Cond 340i (WTW)
Incubator with Timer Rumed (RUBARTH APPARATE)
Oximeter and pH-Meter HQ 40d multi (HACH LANGE)
Piston-stroke pipettes Finnpipette digital and Finnpipette F2 (THERMO SCIENTIFIC)
Spectrophotometer NANOCOLOR® UV/VIS (MACHEREY-NAGEL)
Thermohygrograph THIES
Standard laboratory equipment
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 304 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
The measured concentrations of the test item at the start of the exposure (0 hours) were in the range of 95 to 117% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 96 to 101% of the nominal values.
The measured test item concentrations remained within ± 20% of the nominal concentrations over the test period. This indicates that the test item concentrations were stable during the duration of the test.
Based on the nominal concentrations of the test item sodium p-cumenesulphonate, the 48-hour EC50 for Daphnia magna was > 304 mg/L corresponding to > 121 mg/l active substance.
Results with reference substance (positive control):
The percentage of immobility for the reference item potassium dichromate (SIGMA-ALDRICH, batch
number MKBV0900V, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours.
The EC50 is 2.03 mg/L (validity range 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000)).

Table 1:    EC10-, EC50- (with Confidence Limits) and EC100-Values


(based on the nominal concentrations of the test item)






































Effect levels


 



Test duration


[hours]



sodium p-cumenesulphonate


Nominal test item concentrations


[mg/L]



EC10


(with confidence limits)



24



304



48



297 (134 - > 304)



EC50



24



> 304



48



> 304



EC100



24



> 304



48



> 304



 


Table 3:       Immobilization Rates after 24 and 48 hours of Exposure in the Definitive Test


              (n = 20, divided into 4 replicates with 5 daphnids each)












































































































Nominal test item concentration


[mg/L]



IMMOBILIZATION [%]



24 hours



48 hours



Replicates



Replicates



1



2



3



4



MV



1



2



3



4



MV



304



20



0



20



0



10



20



20



20



0



15



152



0



0



0



0



0



0



0



0



0



0



76.0



0



0



0



40



10



0



0



0



40



10



38.0



0



0



0



0



0



0



0



0



0



0



19.0



0



0



0



0



0



0



0



0



0



0



Control



0



0



0



0



0



0



0



0



0



0



 


Table 4:    Absolute Numbers of immobile daphnids after 24 and 48 hours of Exposure in the Definitive Test


              (n = 20, divided into 4 replicates with 5 daphnids each)












































































































Nominal test item concentration


[mg/L]



Number of immobile Daphnids / Total number of Daphnids



24 hours



48 hours



Replicates



Replicates



1



2



3



4



Sum



1



2



3



4



Sum



304



1 / 5



0 / 5



1 / 5



0 / 5



2 / 20



1 / 5



1 / 5



1 / 5



0 / 5



3 / 20



152



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



76.0



0 / 5



0 / 5



0 / 5



2 / 5



2 / 20



0 / 5



0 / 5



0 / 5



2 / 5



2 / 20



38.0



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



19.0



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



Control



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



0 / 5



0 / 5



0 / 5



0 / 5



0 / 20



 


Table 5:          Measured Concentrations of the Test Item during the Definitive Test




































































Sampling Date



0 hours



48 hours



Nominal concentration


of the test item


[mg/L]



sodium p-cumenesulphonate



Meas. conc.


[mg/L]



%



Meas. conc.


[mg/L]



%



304



288



95



293



96



152



162



107



152



100



76.0



83.1



109



76.5



101



38.0



44.4



117



36.9



97



19.0



20.4



108



18.6



98



Control



< LOQ



< LOQ



QC



5.00



98



4.89



96



Meas. Conc.    = measured concentration of the test item, dilution factor taken into account


%                      = Percent of the nominal test item concentration


LOQ                 = Limit of quantification (5.00 mg test item/L)


QC                    = quality control (5.00 mg test item/L), recovery calculated to nominal concentration


 


 


 


 


The study was performed according to OECD Guideline 202 (2004). The validity criteria were fulfilled.




















Validity criteria (target)ResultValid
The test is unacceptable if more than 10% of the control organisms are immobilized
or show other signs of disease or stress, e.g. discoloration or unusual behavior such
as trapping on the surface of the water, during the 48-hour test period
No daphnidyes
The dissolved O2 concentration in the old media at the end of the exposure should
be ≥ 3 mg/L in all concentration levels and in the control
≥ 7.71 mg/Lyes
Validity criteria fulfilled:
yes
Conclusions:
EC50 > 304 mg/L 48 h corresponding to > 121 mg/l active ingredient
Executive summary:

The short term toxicity to invertebrates of Sodium p-cumenesulphonate was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test. The study was conducted under static conditions over a period of 48 hours with five nominal concentrations of the test item sodium p-cumenesulphonate prepared in a geometric series with a separation factor of 2 (nominal concentrations of 19 to 304 mg/L), corresponding to 7.54 – 121 mg active substance/L.


All test item concentrations and the control were colorless and visually clear throughout the exposure period.


Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control.


The concentrations of the test item sodium p-cumenesulphonate were analytically verified via LCMS/ MS in the fresh media at the start of the exposure (0 hours) and in the old media at the end of the exposure (48 hours) in all concentration levels and in the control.


The measured concentrations of the test item at the start of the exposure (0 hours) were in the range of 95 to 117% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 96 to 101% of the nominal values.


The measured test item concentrations remained within ± 20% of the nominal concentrations over the test period. This indicates that the test item concentrations were stable during the duration of the test. The effect concentrations are based on the analytically verified nominal concentrations of the test item sodium p-cumenesulphonate and the active substance concentrations.


The validity criteria of the test guideline were fulfilled.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
March 26-30, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with no analytical confirmation and some details missing
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: direct dispersion in water
Test organisms (species):
Daphnia magna
Details on test organisms:
no details provided
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
270 mg/L
Test temperature:
21 °C
pH:
7.8 +/- 0.2
Dissolved oxygen:
air saturation
Nominal and measured concentrations:
nominal - 100 mg/L test substance; 65 mg/L active ingredient
no measurements
Details on test conditions:
TEST SYSTEM
- Test vessel: glass
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 250 mL volume filled to 200 mL
- Aeration: no auxiliary aeration
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water - 25 mL of four solutions were added to each litre (final volume) of deionized water. The four solutions were 11.76 g/L of CaCl2.2H2O, 4.93 g/L of MgSO4.7H2O, 2.59 g/L of NaHCO3, and 0.23 g/L KCl.

OTHER TEST CONDITIONS
- Photoperiod: not indicated
- Light intensity: not indicated

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and adverse reactions at 24 hour intervals

TEST CONCENTRATIONS
- Range finding study: yes, but no details provided
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
active ingredient is 65% of test material
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: no adverse reactions (including immobilization)
- Mortality of control: none observed
Results with reference substance (positive control):
no reference substance
Validity criteria fulfilled:
yes
Conclusions:
EC 50 48 h > 100 mg/L corresponding to > 65 mg/L based on active ingredient.
Executive summary:

The short term toxicity to invertebrates of Hydroxybenzene sulphonic acid was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test.
A limit test was performed, exposing daphnids per concentration (in quadruplicate) to a black-control and hydroxyl benzene sulphonic acid of 100 mg/l based on active ingredient. The total test period was 48 hours and met acceptability criteria described in the official protocol. The tested item did not induce acute immobilisation of Daphnia Magna at 100 mg/l after 48 hours of exposure corresponding to 65 mg/l a.i.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read Across from analogue substance
Adequacy of study:
supporting study
Study period:
2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Out of the stock solution 5 concentrations were tested in a geometric series with a dilution factor of 2:
9.50 - 19 - 38.0 - 76.0 - 152 mg test item/L, corresponding to 6.26 - 12.5 - 25 - 50 - 100 mg a.s./L.
- Sampling method: 152 mg test item/L were freshly prepared with dilution water. The dispersion was mixed thoroughly by manual agitation until the solution was visually clear
- Sample storage conditions before analysis: At least 1 g of the test item has been stored at 6 ± 2 °C. Storage at test facility 18 – 25 °C, dark, in the tightly closed original container
Vehicle:
yes
Details on test solutions:
Stock solution, media preparation: 152 mg test item/L were freshly prepared with dilution water. The dispersion was mixed thoroughly by manual agitation until the solution was visually clear.
Preparation of the test item solutions: Out of the stock solution 5 concentrations were tested in a
geometric series with a dilution factor of 2:
9.50 - 19 - 38.0 - 76.0 - 152 mg test item/L, corresponding to
6.26 - 12.5 - 25 - 50 - 100 mg a.s./L.
Control: Dilution water without test item incubated under the same conditions as the test groups
Reference Test : A reference test with potassium dichromate was conducted as an acute immobilization test (acc. to AQS P 9/2 and OECD 202) in Elendt M4 medium under static conditions with a test duration of 24 hours once per month in order to prove the validity of the test system and test conditions at the test facility.
Test vessels: Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses
Test volume: 20 mL
Dilution water: Same composition as the culture medium
Test organisms (species):
Daphnia magna
Details on test organisms:
Test system: Daphnia magna STRAUS, obtained from continuous laboratory cultures.
Reason for the selection of the test system: Daphnia magna is the preferred species in accordance with the test guideline and is bred at the test facility.
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
Breeder: Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
Culture: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, 16 hours illumination; light intensity of max. 1500 lx
Culture medium: Elendt M4, according to OECD 202, Annex 3 (2004), is used.
Feeding of the culture stocks: The daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus. The algae were cultured at the test facility.

Number of daphnids and replicates : 20 daphnids, divided into 4 replicates, each with 5 daphnids, were used for each concentration and the control.
Age of the daphnids at the start of the exposure: Less than 24 hours old daphnids from a healthy stock were used for the study. To achieve this, juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period were used for the test. No first brood progeny was used for the test.
Acclimatization: Acclimatization of the daphnids was not necessary, because the composition of the dilution water was equivalent to the culture medium.

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
see tables attached
pH:
see tables attached
Dissolved oxygen:
see tables attached
Conductivity:
see tables attached
Nominal and measured concentrations:
9.50 - 19 - 38.0 - 76.0 - 152 mg test item/L, corresponding to 6.26 - 12.5 - 25 - 50 - 100 mg a.s./L.
Details on test conditions:
Application: 20 g test solution per replicate were weighed out into each test vessel.
This corresponds to 20 mL per test vessel. The daphnids were inserted with a small amount of dilution water by a pipette.
Test temperature (target): 18 - 22 °C, constant within ± 1 °C
Illumination (target): Diffuse light, light intensity of max. 1500 lx
Photoperiod (target): 16/8 hours light/dark cycle
Feeding: The daphnids were not fed during the study

Type and Frequency of Measurements
Biological Parameters
Immobilization and other observations: Immobilization was determined in all groups after 24 and 48 hours.
An animal was considered immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel.
Observation of other adverse, sub-lethal effects (e.g. discoloration, abnormal behavior etc.) was done, but none occurred during the course of the study.

Water Quality Parameters
Dilution water Prior to the start of the exposure (0 hours), the water quality parameters (i.e. pH-value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured.

Test media
At the start of the exposure (0 hours), the water quality parameters of the fresh media (i.e. pH-value, dissolved oxygen concentration) were measured in one additional replicate (without daphnids) per concentration level and control.
At the end of the exposure (48 hours), the water quality parameters of the old media were measured in one appropriate replicate (containing daphnids) per concentration level and control. The replicate with the highest immobilization rate per concentration level and control was
measured.

Temperature
The incubator temperature was recorded throughout the period of the test.

Quality criteria for the water quality parameters (target)
• The dissolved oxygen concentration in old media should be ≥ 3 mg/L in all concentration levels and in the control.
• The pH should be in the range of 6 – 9.
• The deviation of the final pH-values (old media) from the initial values (fresh media) should not exceed 1.5 units.
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 152 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: corresponding to > 100 mg active substance/L.
Details on results:
The measured concentrations of the test item at the start of the exposure (0 hours) were in the range of 83 to 100% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 89 to 109% of the nominal values.
The measured test item concentrations were within ± 20% of the nominal concentrations over the
test period. This indicates that the test item concentrations were stable during the duration of the
test.
All test item concentrations and the control were colorless and visually clear throughout the exposure period.
The measured water quality parameters (i.e. pH-value, dissolved oxygen concentration, total water hardness and water temperature) were within the acceptable limits. During the test period, the temperature in the incubator was 20.0 – 20.5 °C.
Results with reference substance (positive control):
The percentage of immobility for the reference item potassium dichromate (SIGMA-ALDRICH, batch
number MKBV0900V, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours.
EC50 = 2.03 mg/L (validity range 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000))

The study was performed according to OECD Guideline 202 (2004). The validity criteria were fulfilled:




















Validity criteria (target)ResultValid
The test is unacceptable if more than 10% of the control organisms are immobilized
or show other signs of disease or stress, e.g. discoloration or unusual behavior such
as trapping on the surface of the water, during the 48-hour test period.
No daphnidyes
The dissolved O2 concentration in the old media at the end of the exposure should
be ≥ 3 mg/L in all concentration levels and in the control
≥ 7.60 mg/Lyes
Validity criteria fulfilled:
yes
Conclusions:
EC50 48 h > 152 mg/L corresponding to > 100 mg/L active substance/l
Executive summary:

The short term toxicity to invertebrates of Hydroxybenzene sulphonic acid was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test.


The study was conducted under static conditions over a period of 48 hours with five nominal concentrations of the test item 4 - hydroxybenzenesulphonic acid prepared in a geometric series


with a separation factor of 2 (nominal concentrations of 9.50 to 152 mg/L, corresponding to active substance concentrations of 6.25 to 100 mg active substance/L).


All test item concentrations and the control were colorless and visually clear throughout the exposure period. Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control. The tested item did not induce acute immobilisation of Daphnia Magna at 100 mg/l a.i. after 48 hours of exposure.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from a member of the Hydrotropes category
Adequacy of study:
supporting study
Study period:
July 12-14, 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 60, 120, 250, 500 and 1000 mg/L nominal; all test concentrations were corrected for the 31.2% sample activity. The mean measured concentrations were 39, 150, 220, 470 and 1,020 mg/L.
- Sampling method: from test chambers at 0 and 48 hours of the test. Validated analysis method for recovery of SAR 33-55 in aquatic test water prior to initiation of the test. Results of method validation in report 41658.
- Sample storage conditions before analysis: details on the report
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
A half-liter proportional diluter system described by Mount and Brungs utilizing a Hamilton Micro Lab 420 syringe dispenser, was used for the intermittent introduction of dilution water and tested item into the test chambers
- Evidence of undissolved material (e.g. precipitate, surface film, etc): solutions were clear with no visible floating material
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnid
- Strain: Daphnia magna
- Source: in-house culture maintained since 1977. Original source (strain) was Columbia National Fisheries Research Laboratory
- Age at study initiation (mean and range, SD): <24 hr; first instar
- Weight at study initiation (mean and range, SD): no data
- Feeding during test not fed ACCLIMATION- Acclimation period: from in-house culture of adults monitored for 13-days- Acclimation conditions (same as test or not): same as test
- Type and amount of food: Selenastrum capricornutum and a supplement consisting of fish food and active dry yeast; amount not specified
- Feeding frequency: at least every 3 days
- Health during acclimation (any mortality observed): no mortality and no signs of stress, disease or physical damage
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
152-154 mg/L CaCO3
Test temperature:
19-21 degrees C
pH:
8.4
Dissolved oxygen:
7.4 - 8.2 mg/L (85 - 94% saturation)
Salinity:
freshwater
Nominal and measured concentrations:
0, 60, 120, 250, 500 and 1000 mg/L nominal; 0, 39, 150, 220, 470 and 1020 mg/L measured
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 1 L glass beaker
- Aeration: flow through- Type of flow-through (e.g. peristaltic or proportional diluter): proportional diluter
- Renewal rate of test solution (frequency/flow rate): 3.9 mL/min (5.6 replacement volumes in 24 hours)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Loading rate: 1 daphnid per 100 mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: hard blended lab water
- Total organic carbon: <1 mg/L- Particulate matter: 0 - 0.2 mg/L
- Metals, Pesticides: checked
- Alkalinity: 152-154 mg/L
- Conductivity: 320-340
- Culture medium different from test medium: no difference
- Intervals of water quality measurement: time 0 and 48 hours

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted
- Photoperiod: 16 light / 8 dark with 30-minute transition periods
- Light intensity: 661-687 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, immobility, surfacing, erratic movement and daphnids on the bottom; observations at 24 hour intervals

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2X
- Range finding study: yes
- Test concentrations DRF: 1, 10, 100, 1000 and 10,000 mg/L nominal
- Results used to determine the conditions for the definitive study: 1 daphnid dead at 1000; all dead at 10000
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
470 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 1 020 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mobility
Details on results:
Three and five daphnids were on the bottom of the test chambers in both replicates at the 1020 mg/L exposure. One daphnid was immobile in the control, one at 39 mg/L and one at 470 mg/L. The NOEC was determined to be 470 mg/L.
Validity criteria fulfilled:
yes
Conclusions:
The 48-hr EC50 was >1020 mg/L
The NOEC was 470 mg/L.
Executive summary:

The short term toxicity of Calcium Xylenesulphonate to Daphnia magna was assessed following EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids). Water quality parameters of temperature, dissolved oxygen, and pH were measured throughout the test and were within acceptable limits. The daphnids used for this study were f"rrst-instar daphnids « 24 hours old) and were cultured at the lab. Culture records indicated the adult daphnids were in good condition.
The test concentrations of the tested item were analytically quantified at 0 and 48 hours. The study was conducted at the mean measured concentrations of 39, 150, 220, 470 and 1,020 mg/L. Ten daphnids were added to each replicate beaker for a total of 20 daphnids per test concentration and control. The 24- and 48-hour ECso values were both > 1020 mg/L. The results indicated the 48-hour no-observed effect concentration (NOEC) was 470 mg/L, based on the lack of immobility or significant abnormal effects at this concentration.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from analogue substance
Adequacy of study:
supporting study
Study period:
2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well described GLP compliant study conducted to recognized international test guidelines
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Sampling times were a 0h and 48h
Volume: 1 ml
Samples were stored in a freezer until analysis
Vehicle:
no
Details on test solutions:
Stock solution of 100 mg/L in ISO-medium stired for 5-8 minutes
The final test solutions were all clear and colourless.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnid
- Strain: not indicated
- Source: in-house culture with a known history
- Age at study initiation (mean and range, SD): <24h
- Feeding during test: no

ACCLIMATION
- Acclimation period: in house culture
- Acclimation conditions (same as test or not): yes
- Type and amount of food: suspension of freshwater algae
- Feeding frequency: daily
- Health during acclimation (any mortality observed): not indicated

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
250 mg/L as CaCO3
Test temperature:
19.1-19.7 C
pH:
control - 7.9-8.0
100 mg/L - 5.9-7.2
Dissolved oxygen:
9.2-9.7 mg/L
Nominal and measured concentrations:
nominal - 0 and 100 mg/L
measured - 0 and 103 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 100 mL with 80 mL solution
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: not indicated

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO-medium with hardness of 250 mg/L as CaCO3 and pH of 8.0 +/- 0.2
- Culture medium different from test medium: no
- Intervals of water quality measurement: 0h and 48h except temperature which is continuously monitored

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light
- Light intensity: not indicated

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility including mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: limit test
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 103 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
other: immobility including mortality
Details on results:
No immobilization or mortality at 103 mg/L
The pH of the solution at 103 mg/L was initially 5.9 (which is just below the 6.0 guideline limit) at the 0h but increased to 7.2 at 48h. This was not considered to have an effect on the daphnids.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Mortality: no mortality at 0, 0.1, 0.18 or 0.32 mg/L; 5% at 0.56 mg/L, and 100% at 1.0 and 1.8 mg/L
- EC50/LC50: appx 0.75 mg/L
- Other:
Validity criteria fulfilled:
yes
Conclusions:
EC50 > 100 mg/l based on active ingredient
Executive summary:

The short term toxicity to invertebrates of Benzene sulphonic acid was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test. A limit test was performed, exposing 20 daphnids per concentration (in quadruplicate, 5 per vessels) to a black-control and a benzene sulphonic acid of 100 mg/l based on active ingredient. The total test period was 48 hours and met acceptability criteria described in the official protocol. The tested item did not induce acute immobilisation of Daphnia Magna at 100 mg/l a.i. after 48 hours of exposure.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
other: Read across from analogue substance
Adequacy of study:
supporting study
Study period:
March 24-26, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with no analytical confirmation and some details missing
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: direct dispersion in water
Test organisms (species):
Daphnia magna
Details on test organisms:
no details provided
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
270 mg/L
Test temperature:
21C
pH:
7.8 +/- 0.2
Dissolved oxygen:
air saturation
Nominal and measured concentrations:
nominal - 100 mg/L test substance; 62.8 mg/L active ingredient
no measurements
Details on test conditions:
TEST SYSTEM
- Test vessel: glass
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 250 mL volume filled to 200 mL
- Aeration: no auxiliary aeration
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water - 25 mL of four solutions were added to each litre (final volume) of deionized water. The four solutions were 11.76 g/L of CaCl2.2H2O, 4.93 g/L of MgSO4.7H2O, 2.59 g/L of NaHCO3, and 0.23 g/L KCl.

OTHER TEST CONDITIONS
- Photoperiod: not indicated
- Light intensity: not indicated

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and adverse reactions at 24 hour intervals

TEST CONCENTRATIONS
- Range finding study: yes, but no details provided
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
active ingredient is 62.8% of test material
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: no adverse reactions (including immobilization)
- Mortality of control: none observed
Results with reference substance (positive control):
no reference substance
Validity criteria fulfilled:
yes
Conclusions:
EC 50 48 h > 100 mg/L correspondign to > 62.8 mg/L based on active ingredient.
Executive summary:

The short term toxicity to invertebrates of Benzene sulphonic acid was assessed according to official guideline OECD 202, Daphnia sp. Acute Immobilisation Test. A limit test was performed, exposing daphnids per concentration (in quadruplicate) to a black-control and a Benzene sulphonic acid of 100 mg/l based on active ingredient. The total test period was 48 hours and met acceptability criteria described in the official protocol. The tested item did not induce acute immobilisation of Daphnia Magna at 100 mg/l a.i. after 48 hours of exposure.

Description of key information

EC50 for freshwater invertrebrates = 100 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Dose descriptor:
EC50
Effect concentration:
100 mg/L

Additional information

The ASA Category comprises the following 5 aromatic sulphonic acids:


TSA, Toluene-4-sulphonic acid (EC 203-180-0, CAS 104-15-4)


XSA, (Xylenes and 4-ethylbenzene) sulphonic acid (EC 701-247-3, CAS -) Former EC 246-839-8


CSA, p-cumene sulphonic acid (EC 240-210-1, CAS 16066-35-6)


BSA, Benzene sulphonic acid (EC 202-638-7, CAS 98-11-3)


HBSA, 4-hydroxybenzensulphonic acid (EC No. 202-691-6, CAS No. 98-67-9)


The short-term toxicity on aquatic invertebrates was assessed with available studies on ASA and related salt (Hydrotropes category). Hydrotropes are completely dissociated in water and could be used in read across for the related acid form. The Short-term toxicity testing on invertebrates has been tested on BSA, TSA and HBSA. The endpoint for XSA and CSA was assessed with the tests performed on the related salt.


The key study is a 2005 Guideline (OECD 202) study with a 99.8% analytically pure benzenesulfonic acid (CAS No. 98-11-3) conducted under GLPs. It is a well documented limit test with analytical confirmation. There was no mortality or immobility observed at 100 mg/L (nominal) / 103 mg/L measured. The test conditions were confirmed to be valid and the reference substance produced expected results. The 48h-EC50 for 98-11-3 is reported as >100 mg/L.


The 2005 study with benzenesulfonic acid is the only one of the three available studies on BSA and TSA with sufficient details and adherence to OECD guidelines to use to establish the 48h-EC50 of >100 mg/L. The other 2 studies give support to this endpoint value.


There is a 1995 supporting study conducted following OECD 202 but without analytical confirmation and no confirmation of GLP compliance. This limit test was conducted with a 65% pure (in water) benzenesulfonic acid (CAS No, 98-11-3). There was no toxicity observed and the 48h-EC50 is reported as >100 mg/L.


There is also a 1983 study of p-toluenesulfonic acid (CAS No. 104-15-4) designated a "weight-of-evidence" study. There were multiple, nominal test concentrations (5000, 1000, 100, 10 and 0), however, the exposure duration was 24 hours instead of the required 48 hours. There was no toxicity in the 0 and 10 mg/L exposures, 5% at 100 mg/L, 35% at 1000 mg/L and 10% at 5000 mg/L. There are insufficient details and no plausible explanation for the lower toxicity at 5000 mg/L. The results indicate the 24h-EC50 is above 1000 mg/L.


In 2020 HBSA was tested in GLP according to OECD 202 under static conditions over a period of 48 hours with five nominal concentrations of the test item prepared in a geometric series.
Based on the nominal concentrations of the test item 4 - hydroxybenzenesulphonic acid, the 48-hour EC50 for Daphnia magna was > 152 mg/L, corresponding to > 100 mg active substance/L.


Further studies are available for the sodium salt of Toluene, Xylene and Cumene derivatives (Hydrotropes) and in all cases the EC50 48 h was higher than 100 mg/L.


It can be concluded that for all the members of the category the representative result for EC50 48h on daphnia magna is > 100 mg/L