Benzenesulfonic acid, 4-C20-24 (even numbered) sec-alkyl derivs., sodium salts

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 7, 1984-June 22, 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study done according to national guideline. (Study done before implementation of GLP, but adhered to GLP comparable quality conditions.

Qualifier:

according to guideline

Guideline:

other: EPA-600/9-78-018

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

not specified

Details on sampling:

no information

Details on test solutions:

A stock solution of 1,000 mg product/1000 ml DI water was prepared. From this, a second stock solution was prepared with a dilution factor of 20.

Test organisms (species):

other: Chlorella kessleri

Test type:

static

Water media type:

freshwater

Total exposure duration:

15 d

Nominal and measured concentrations:

0, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, 1000 mg/L

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: 100 ml DI water, and 10 ml mineral medium
- Initial cells density: 10^4 cells/ml
- Control end cells density: 3.7 x 10^15
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water, AAP-Medium


OTHER TEST CONDITIONS
- Light intensity and quality: ~2000 lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : number of cells was determined on days 5, 7, 12, and 15

Duration:

15 d

Dose descriptor:

NOEC

Effect conc.:

3.1 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

cell number

Duration:

15 d

Dose descriptor:

LOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

cell number

Number of cells/ml (average of 3 replicates)

Concentration (mg/L)	5 days	7 days	12 days	15 days
0	7.2 x 10^5	1.5 x 10^6	3.9 x 10^6	3.7 x 10^6
0.01	5.9 x 10^5	1.4 x 10^6	2.4 x 10^6	3.2 x 10^6
0.03	3.0 x 10^5	1.4 x 10^6	3.4 x 10^6	3.6 x 10^6
0.1	2.6 x 10^5	1.3 x 10^6	2.4 x 10^6	3.0 x 10^6
0.3	2.5 x 10^5	1.3 x 10^6	3.2 x 10^6	3.5 x 10^6
1	2.6 x 10^5	1.3 x 10^6	2.5 x 10^6	2.8 x 10^6
3	1.6 x 10^5	1.0 x 10^6	2.3 x 10^6	3.2 x 10^6
10	3.9 x 10^5	6.3 x 10^5	1.5 x 10^6	1.7 x 10^6
30	1.9 x 10^5	3.9 x 10^5	1.1 x 10^6	1.4 x 10^6
100	2.9 x 10^4	4.5 x 10^4	4.2 x 10^4	6.1 x 10^4
300	1.3 x 10^4	2.7 x 10^4	1.9 x 10^4	1.1 x 10^4
1000	1.6 x 10^4	2.6 x 10^4	2.5 x 10^4	2.0 x 10^4

Conclusions:

The NOEC for algae was 3.1 mg/L (active ingredient), and the LOEC was 10 mg/L (active ingredient).

Executive summary:

Chlorella kessleri algae was exposed to concentrations of 0, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, and 1000 mg/L (nominal) of test substance for 15 days. The number of cells were counted to determine the toxicity of the test substance. The NOEC was 3.1 mg/L active ingredient, and the LOEC was 10 mg/L active ingredient. The data are for C11.6 LABS Na and no normalization is required.

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Dec. 8, 1992-Dec. 11, 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Qualifier:

according to guideline

Guideline:

other: 88/302/EWG

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

TEST ORGANISM
- Source (laboratory, culture collection): Institut fur Wasser-, Boden- und Lufthygiene, Berlin

ACCLIMATION
- Acclimation period: 3 days

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Test temperature:

24 ± 2 °C

pH:

7.7-9.0

Nominal and measured concentrations:

0, 0.6, 2.4, 10, 40, and 160 mg/L

Details on test conditions:

TEST SYSTEM
- Initial cells density: 2 x 10^4 cells/ml
- Control end cells density: 95 x 10^4 cells/ml
- No. of vessels per concentration (replicates): 8
- No. of vessels per control (replicates): 8

OTHER TEST CONDITIONS
- Light intensity and quality: 8000 Lux

Duration:

72 h

Dose descriptor:

other: EbC50

Effect conc.:

47.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

72 h

Dose descriptor:

other: ErC50

Effect conc.:

127.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

2.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

The pH as measured at the end of the experiment was very high (pH=9.0), however, as growth was not affect, the experiment was considered valid.

Cell count ( x 10⁴cells/ml)

Concentration (mg/L)	0 hrs	24 hrs	48 hrs	72 hrs
Control	2	8	29	95
0.6	2	8	30	89
2.4	2	9	28	85
10	2	9	27	75
40	2	8	20	48
160	2	7	9	8

Validity criteria fulfilled:

yes

Conclusions:

The 72-hr NOEC for algae was 2.4 mg/L. The 72-hr EbC50 was 47.3 mg/L, and the 72-hr ErC50 was 127.9 mg/L for algae.

Executive summary:

Scenedesmus subspicatus was exposed to concentrations of 0, 0.6, 2.4, 10, 40, or 160 mg/L of test substance for 72 hrs. The cell concentration was measured at 24, 48, and 72 hrs. The 72-hr NOEC for algae was 2.4 mg/L. The 72-hr EbC50 was 47.3 mg/L, and the 72-hr ErC50 was 127.9 mg/L for algae. The NOEC of 2.4 mg/L does not require normalization since the compound tested was C11.6 LABS Na.

Reference 3

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982-09-17 To 1982-10-01

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: Payne AG, Hall RH, 1979, in Aquatic Toxicology, ASTM, STP 667, pp. 171-180; and US EPA, 1978, The Selenastrum capricornutum Printz algal assay. Deviations, reliability, and validity will be evaluated using the USEPA OPPTS 850.5400 (1996) guideline.

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

not specified

Remarks:

(pre-GLPs)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Analytical monitoring:

no

Details on sampling:

- Concentrations: Not available
- Sampling method: Not available
- Sample storage conditions before analysis: Not available

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A primary stock solution of X0235.01 was prepared by adding a weighed amount of test material to algal growth medium and other stock solutions were prepared by serial dilution. A growth medium control was conducted concurrently and contained no test material. All test concentrations, except 1,000 ppm, were prepared by pipeting 1 milliliter (m2) of the appropriate stock solution to each test flask. The 1,000 ppm test concentration was prepared by adding 50ml of primary stock solution to each test flask.
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: AAP medium
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Not available
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms (species):

Microcystis aeruginosa

Details on test organisms:

TEST ORGANISM
- Common name: Green algae
- Strain: Not available
- Source (laboratory, culture collection): University of Texas at Austin, Austin, Texas
- Age of inoculum (at test initiation): Not available
- Method of cultivation:


ACCLIMATION
- Acclimation period: Not available
- Culturing media and conditions (same as test or not): Same as test
- Any deformed or abnormal cells observed: Not available

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

120 h

Remarks on exposure duration:

96 hour results reported here

Post exposure observation period:

Yes (9 days)

Hardness:

Not available

Test temperature:

Static bioassays were conducted at 24 ± 1 °C in water bath

pH:

Not available

Dissolved oxygen:

Not available

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal laboratory exposure concentrations based on active ingredient were control, 0.01, 0.05, 0.1, 0.5, 1.0 and 1,000mg/L
No measurements of test concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed: Not available
- Material, size, headspace, fill volume: Test containers were 125-ml flasks with 50 ml of test medium
- Aeration: Not available
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Not available
- Initial cells density: 1x10(5) cells/ml
- Control end cells density: 18.5x10(5) cells/ml
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3


GROWTH MEDIUM
- Standard medium used: Yes, AAP nutrient medium


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Regular Algal Assay Procedure (AAP) medium prepared with deionized water
- Total organic carbon: Not available
- Particulate matter: Not available
- Metals: Not available
- Pesticides: Not available
- Chlorine: Not available
- Alkalinity: Not available
- Ca/mg ratio: Not available
- Conductivity: Not available
- Culture medium different from test medium: No
- Intervals of water quality measurement: Not available

OTHER TEST CONDITIONS
- Sterile test conditions: Yes
- Adjustment of pH: Not applicable
- Photoperiod: Not available
- Light intensity and quality: Approximately 2,000 lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Hemacytometer
- Cell counts were made on day 0,2,3,4,5.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2-5 (except for the highest test concentration of 1000 mg/L)
- Justification for using less concentrations than requested by guideline: Not applicable
- Range finding study: no

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

0.91 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

cell number

Remarks on result:

other: 95%CL: 0.84- 10mg/L

Details on results:

- Exponential growth in the control (for algal test): Yes (18.5x growth)
- Observation of abnormalities (for algal test): Not available
- Any stimulation of growth found in any treatment: Not available
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: None

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The EC50 values were calculated using the following three statistical methods: moving average angle analysis, Probit analysis, and binominal probability. The results of the moving average angle method were used to report the EC50 values.

Comparison was also made to in situ studies conducted in which lake water was bottled and suspended in Lake Acton (Ohio) for 3 hour periods. The mean 3-h EC₅₀(photosynthesis) for the in situ studies was 3.4 mg/L (0.5-8.0 mg/L).Using the acute to chronic ratio calculation (documented in Annex 3 of the LABS Na SIAR), the EC₅₀/3 for Microcystis is 0.3 mg/L. The NOEC normalized by van de Plassche et al. (1999) to C_11.6 LABS Na was 0.35 mg/L. 

Validity criteria fulfilled:

yes

Conclusions:

In a 96 hour algae growth inhibition test, the EC50 of LABS Na to Microcystis aeruginosa was 0.91 mg/L.

Executive summary:

In a 96 hour algae growth inhibition test, the green algae Microcystis aeruginosa was exposed to Linear Alkylbenzene Sulfonate (C11.9 LABS Na) in accordance with USEPA OPPTS 850.5400. The nominal test concentrations were 0 (control), 0.01, 0.05, 0.1, 0.5, 1.0 and 1,000 mg a.i./L.

The 96 hour EC50, based on cell number, was 0.91 mg a.i./L (0.84 -10.0). The NOEC is estimated to be 0.35 mg/L for C11.6 LABS Na (van de Plassche et al., 1999).

This algal growth inhibition test satisfies the guideline requirements of the USEPA OPPTS 850.5400.

Reference 4

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Aug. 20, 1982-Sept. 13, 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The studies are very well documented in this peer-reviewed publication and deserving of a reliability 2.

Qualifier:

according to guideline

Guideline:

other: Payne AG, Hall RH, 1979, in Aquatic Toxicology, ASTM, STP 667, pp. 171-180; and US EPA, 1978, The Selenastrum capricornutum Printz algal assay. Deviations, reliability, and validity will be evaluated using the USEPA OPPTS 850.5400 (1996) guideline.

Deviations:

no

Principles of method if other than guideline:

Standard Practice for Conducting Toxicity Tests with Microalgae, Draft #7, Philadelphia, PA.
Assumed 1 x 10^4 cells/mL because ASTM protocol, but not reported.

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A primary stock solution of X0235.01 was prepared by adding a weighed amount of test material to algal growth medium and other stock solutions were prepared by serial dilution. A growth medium control was conducted concurrently and contained no test material. All test concentrations, except 1,000 ppm, were prepared by pipeting 1.0 milliliter (ml) of the appropriate stock solution to each test flask.
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: AAP medium
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Not available
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms (species):

Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Green algae
- Strain: Not available
- Source (laboratory, culture collection): University of Texas at Austin, Austin, Texas
- Age of inoculum (at test initiation): Not available
- Method of cultivation: Not available

ACCLIMATION
- Acclimation period: Not available
- Culturing media and conditions (same as test or not): Same as test
- Any deformed or abnormal cells observed: Not available

Test type:

other: static open-system

Water media type:

not specified

Limit test:

no

Total exposure duration:

96 h

Hardness:

Total mean water hardness was 137 mg/L as CaCO3.

Test temperature:

Mean test temperature was 23.6 (21.2-25.6 °C).

pH:

The pH range was 6.8 to 7.2.

Dissolved oxygen:

Mean dissolved oxygen was 9.1 mg/L.

Nominal and measured concentrations:

Nominal: 0, 16, 32, 63, 125, 250, 500, and 1000 mg/L first test
0.1, 1.0, 5.0, and 10 mg/L second test

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed: Not available
- Material, size, headspace, fill volume: Test containers were 125-ml flasks with 50 ml solution
- Aeration: Not available
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Not available
- Initial cells density: Both test 1 and test 2: 2x10(4) cells/ml
- Control end cells density: Test 1: 318x10(4); Test 2: 245x10(4)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3


GROWTH MEDIUM
- Standard medium used: Yes, AAP nutrient medium


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: AAP media prepared with deionized water
- Total organic carbon: Not available
- Particulate matter: Not available
- Metals: Not available
- Pesticides: Not available
- Chlorine: Not available
- Alkalinity: Not available
- Ca/mg ratio: Not available
- Conductivity: Not available
- Culture medium different from test medium: No
- Intervals of water quality measurement: Not available


OTHER TEST CONDITIONS
- Sterile test conditions: yes/no Not available
- Adjustment of pH: Not applicable
- Photoperiod: Not available
- Light intensity and quality: Approximately 2,000 lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Hemacytometer
- Cell counts were made on day 0,2,3,4,5.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 (test 1). 2-10 (test 2).
- Justification for using less concentrations than requested by guideline: Not applicable
- Range finding study: no

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

29 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: cell density

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: cell density

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: cell density

Details on results:

- Exponential growth in the control (for algal test): Yes. Test 1 (159x growth). Test 2 (122x growth)
- Observation of abnormalities (for algal test): In all test concentrations >=1.0 mg/L, the algal cells were abnormally large and malformed. Because of this observation, algae previously exposed to 1000 mg/L were tested for their ability to recover. Their growth was similar to the growth of the control during a 9-day recovery period in test material-free medium, and the algal cells had returned to normal appearance.
- Any stimulation of growth found in any treatment: Not available
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: None

Results of the laboratory studies were compared with enclosure studies conducted with natural phytoplankton assemblages. Concentrations in the enclosures that first altered community structure were found to be between 27 and 108 mg/L. 

Validity criteria fulfilled:

yes

Conclusions:

The 96-hr EC50 for aquatic algae was 29.0 mg/L based on cell density.

Executive summary:

This study was done to determine the toxicity of the test substance to aquatic algae. Selenastrum capricornutum was exposed to the test substance for 96 hrs. During this time, the cell density of the algae was measured. The 96 -hr NOEC was 0.5 mg/L. The 96 -hr LOEC was 1 mg/L. The 96 -hr EC50 was 29.0 mg/L based on cell density. Normalized to C11.6 LABS Na, the value is 0.58 mg/L for the NOEC (method: van de Plassche et al., 1999).

Description of key information

No study was available on the toxicity of the test substance to aquatic algae and cyanobacteria. Therefore, read across is performed using four studies from the related substance benzenesulfonic acid (LABS Na). The study of Muehlberg et al. (1984) obtained a 15 day NOEC of 3.1 mg/L and 15 day LOEC of 10 mg/L for the algae Chlorella kessleri. Ward et al. (1982) obtained a 96 hour EC50 of 0.91 mg/L for the cyanobacterium Microcystis aeruginosa. Lewis al. (1986) obtained a 96 hour EC50 of 29 mg/L and a 96 hour NOEC of 0.5 mg/L for the algae Pseudokirchneriella subcapitata. Scholz al. (1992) obtained a 72 hour ErC50 of 127.9 mg/L and 72 hour NOEC of 2.4 mg/L for the algae Desmodesmus subspicatus. A worst case approach was applied for chemical safety assessment and the most conservative (no-)effect concentrations were used (96 hour EC50 of 0.91 mg/Lfor Microcystis aeruginosa and 96 hour NOEC of 0.5 mg/L for Pseudokirchneriella subcapitata).

Key value for chemical safety assessment

EC50 for freshwater algae:

0.91 mg/L

EC10 or NOEC for freshwater algae:

0.5 mg/L