Ammonium (xylenes and 4-ethylbenzene)sulfonates

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from another member of the category

Adequacy of study:

key 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.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from another member of the 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:

EC50 is > 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) sulfonatese 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.).

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from another member of the 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)	Result	Valid
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 daphnid	yes
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/L	yes

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.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from another member of the 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 cprresponding 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.).

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: Read across from another member of the 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.

Description of key information

The HYDROTOPES Category comprises the following 6 substances:
STS - Sodium toluene 4-sulphonate (CAS 657-84-1, EC 211-522-5)
SXS - Sodium (xylenes and 4-ethylbenzene) sulphonate (EC 701-037-1)
NH4XS - Ammonium (xylenes and 4-ethylbenzene) sulphonate (EC 943-024-5)
SCS - Sodium p-cumenesulphonate (CAS 15763-76-5, EC 239-854-6)
KCS - Potassium p-cumenesulphonate (CAS 164524-02-1, EC 629-764-9)
NH4CS - Ammonium p-cumenesulphonate (CAS 680972-33-2, EC 811-484-5) 
In addition CaXS (Calcium Xylenesulphonate, CAS 28088-63-3, EC 248-829-9) was evaluated for complete the assessment despite it is not registered under REACH.

There are six available studies on short term toxicity to daphnia performed on the three subgroups (toluene, xylene and cumene).
The results of the available studies are:
STS (2020, GLP): EC50 = 217 mg/l (a.i.)
STS (1995, GLP): EC50 = 54 mg/l (disregarded)
SXS (1993, no GLP): EC50 > 1000 mg/l
SXS (1995, GLP not specified): EC50 > 40.3 mg/l (a.i.) Limit test, highest concentration tested
CaXS (1994, GLP): EC50 > 1020 mg/l (a.i.)
SCS (1995, GLP not specified): EC50 > 40.3 (a.i.)
SCS (2020, GLP): EC50 > 121 mg/l (a.i.)
The key study used for the assessment is the test performed on SCS (2020, GLP) with the 48-hour EC50 value > 121 mg/L.

The other availbale studies are:

- SXS: a 1993 USEPA Guideline study (EPAOTS 797.1300) Aquatic Invertebrate Acute Toxicity. The study was not conducted following GLP principles but there is considerable detail provided. Daphnids were exposed to five nominal test concentrations of sodium xylenesulphonate and immobility was monitored for 48 hours. The exposure was static and there was no analytical measurement. No immobilization was observed in the test concentrations and the nominal 48-hour EC50 was >1000 mg/L active substance.

- CaXS: a 1994 study where the 48 hour EC50 was 1020 mg/L measured active substance based on immobilization of 30% and 50% of the organisms in each of two replicate exposures at this concentration. The flow-through study (Ruetgers-Nease Corporation, 1994) with analytical measurements was conducted with calcium xylenesulphonate. The NOEC was 470 mg/L measured active substance.

- SXS: a 1995 OECD 202 Guideline supporting study with the test substance sodium xylenesulphonate that reported no immobility of daphnids at any concentration up to the highest concentration tested (100 mg/L); corresponding to an EC50 of > 40.3 mg/L active substance.

- SCS: a 1995 OECD 202 Guideline supporting study 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.).

- STS: a 1995 EU Method C.2 study of sodium toluenesulphonate conducted under GLP requirements followed a static test method. The EC50 equal to 54 mg/l. This study result is considered to be not reliable due to the missing information about impurities considering that a new study on STS were performed in 2020 with a EC50 > 100 mg/l based on active ingredient. The study result is considered to be incosistent and is not taken into account in PNEC derivation.

 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

121 mg/L