Sodium 4-vinylbenzenesulphonate

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

January 4, 1999 to March 30, 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None specified

Analytical monitoring:

yes

Details on sampling:

10 ml of test samples were taken from each test group at the start of test and at 72 hours after start and analyzed with HPLC. At the analysis of test solution, standard solution was also measured for each test sample analysis and concentrations of test material were quantitatively determined with peak height of test material.

Vehicle:

no

Details on test solutions:

1000 mg/L of mother test solution was prepared by adding sodium p-toluenesulfonate to test medium. This solution was added into medium sterilized with autoclave to prepare test solution for each concentration group.

Test organisms (species):

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

Details on test organisms:

Single-cell green algae of Selenastrum capricornutum is used for the test.
This species is aseptically continuous cultured in this laboratory. Growth Inhibition concentration (EbC50) at 72 hours for standard substance (potassium dichromate, special grade reagent) was 0.84 mg/L
Preculture Cultivation
Algae cultured for 4 days before the start of exposure under the same conditions of the test were used for the test.

Test type:

not specified

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Post exposure observation period:

No post exposure observation period specified in the study report.

Hardness:

Not specfied in the study report.

Test temperature:

Temperature inside algae incubation test equipment was constantly 23.1 °C for 72 hours during exposure period.

pH:

pHs of the test solution were 8.3-8.6 at the start of exposure and 9.1- 10.1 at the end of exposure.

Dissolved oxygen:

Not specified in the study report.

Salinity:

Not specified in the study report.

Nominal and measured concentrations:

Nominal concentrations of 1000, 320, 100, 32, 10, 3.2 and 1.0 mg/L of test material groups and control group were set up.

Details on test conditions:

Test Method
Test conditions
1) Cultivation method: Shaking cultivation (100 rpm)
2) Temperature: 23±2°C
3) Exposure period: 72 hours
4) Volume of test solution: 100 mL (OECD medium)
5) Lighting: 4000-5000 lux (Continuous lighting)
6) Initial cell concentration: 1 X 104 cells/mL

Growth medium
Growth medium described in OECD Guidelines for the testing chemicals was used in both preliminary cultivation and test.

Test vessels. Algae culturing test equipment and instruments
Test vessels: 300mL Erlenmeyer flask made of glass
Algae culturing test equipment: Algae Test incubator (Miyamoto Riken Kogo GT-40S)
Optical microscope: Olympus inverted incubation microscope (IMT413)
Cell counter: Counting chamber
pH meter: Toa Electronics (HM-30V)
Thermometer: Glass mercury thermometer
Illuminometer: Sibata Scientific Technology Ltd. (AN-F9)

Test procedure
After counting the number of algae cells incubated in the preculture, a certain amount of incubated solution obtained in preliminary cultivation was added into the test vessels containing test solution to adjust cell concentration in the test solution to be 1 X 104 cells/ml.
Each test solution was set up in the incubation equipment at 23 ± 2°C and started test. Cell concentrations were counted at 24, 48 and 72 hours. Cell concentrations were measured by taking 2.0 ml of test solution from each test vessel and counting the number of the cells on the counting chamber using microscope.
pH of the test solutions were measured for 3 replicates and a backup. These value were assigned as pH at the start of exposure. At the end of test, pH for all 3 replicates were measured. During test period, temperature and lighting intensity were measured at least once a day.

Reference substance (positive control):

no

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

Environmental factors which might affect reliability of the test result.
None

Concentration of test material in the test solution.
Concentrations of test material at the start of exposure were from 1.0 to 1010 mg/L (nominal concentration from 1.0 to 1000 mg/L). After 72 hours, the concentrations of test material were from 1.0 to 1030 mg/L.

Algae growth curve
Cell concentration of the control increased more than 80 times after 72 hour-incubation. This result showed that cells growth was normal under test conditions.

Fifty percent effective concentration (EC50) and No Observed Effect Concentration (NOEC)
1) Growth inhibition concentration (EbC50) by comparison of area under growth curve.
EbC50 (0-72h) was more than 1000 mg/L. The highest concentration where no significant difference was observed when compared with control (No Observed Effect Concentration (NOECb)) was 10 mg/L.
2) Growth inhibition concentration (ErC50) by comparison of growth rate
ErC50 (24-48 h) and ErC50 (24-72h) were more than 1000 mg/L respectively.
The highest concentration where no significant difference was observed when compared with control (No Observed Effect Concentration (NOECr)) was 1000 mg/L.

Temperature and pH
Temperature inside algae incubation test equipment was constantly 23.1°C for 72 hours during exposure period. pHs of the test solution were 8.3-8.6 at the start of exposure and 9.1- 10.1 at the end of exposure.

Results with reference substance (positive control):

Reference substance not utilised in this study.

Reported statistics and error estimates:

None detailed in the study report.

Measured Concentrations During a 72-Hour Exposure to Selenastrum capricornutum

Nominal Concentration mg/L	Measured Concentration (mg/L)
	0 Hour mg/L	Percent of Nominal	72 Hour mg/L	Percent of Nominal
1000	1010	101	1030	103
320	327	102	328	103
100	101	101	104	104
32	32.9	103	33.1	103
10	10.1	101	10.4	104
3.2	3.3	103	3.4	106
1.0	1.0	100	1.0	100
Control	N.D.	-	N.D.	-

 

Cell Density of Selenastrum capricornutum

Nominal Concentration mg/L	No.	Cell Density (x10^4cells/mL)
		0 Hour	24 Hour	48 Hour	72 Hour
Control	1 2 3	1.00 1.00 1.00	6.00 6.25 6.50	28.25 26.5 27.75	94.25 91.75 89.75
	Average S.D.	1.0	6.25 0.25	27.50 0.90	91.92 2.25
1.9	1 2 3	1.00 1.00 1.00	6.75 6.00 5.50	28.50 27.75 26.00	84.75 81.50 87.75
	Average S.D.	1.00	6.08 0.63	27.42 1.28	84.67 3.13
3.2	1 2 3	1.00 1.00 1.00	6.00 7.00 6.25	30.00 26.75 28.75	86.00 84.25 85.25
	Average S.D.	1.00	6.42 0.52	28.50 1.64	85.17 0.88
10	1 2 3	1.00 1.00 1.00	5.50 6.25 5.75	27.25 28.50 23.50	83.00 88.25 87.25
	Average S.D.	1.00	5.83 0.38	26.42 2.60	86.17 2.79
32	1 2 3	1.00 1.00 1.00	6.00 5.25 5.25	26.25 23.75 23.00	78.50 80.00 85.50
	Average S.D.	1.00	5.50 0.43	24.33 1.70	81.33 3.69
100	1 2 3	1.00 1.00 1.00	5.75 5.25 5.50	27.50 25.25 23.25	75.50 79.50 77.00
	Average S.D.	1.00	5.50 0.25	25.33 2.13	77.33 2.02
320	1 2 3	1.00 1.00 1.00	4.50 5.25 4.00	24.25 24.50 24.50	69.50 80.00 71.50
	Average S.D.	1.00	4.58 0.63	24.42 0.14	73.67 5.58
1000	1 2 3	1.00 1.00 1.00	4.50 4.25 4.25	22.75 25.75 25.75	74.25 72.75 77.00
	Average S.D.	1.00	4.33 0.14	24.75 1.73	74.67 2.16

 

Growth Inhibition of Selenastrum capricornutum

Concentration mg/L		Area A(0-72h)	Inhibition (%)	Rate	Inhibition (%)	Rate	Inhibition (%)
			IA(0-72h)	μ (24-48)	Im (24-48)	μ (24-)	Im(24-72)
Control	1	1893	-2.2	0.0646	-4.7	0.0574	-2.5
	2	1827	1.4	0.0602	2.4	0.0560	0.0
	3	1839	0.8	0.0605	1.9	0.0547	2.3
	Average	1853	0.0	0.0617	0.0	0.0560	0.0
1.0	1	1803	2.7	0.0600	2.8	0.0527	5.9
	2	1728	6.7	0.0638	-3.4	0.0544	2.9
	3	1749	5.6	0.0647	-4.9	0.0577	-3.0
	Average	1760	5.0	0.0628	-1.8	0.0549	2.0
3.2	1	1836	0.9	0.0671	-8.8	0.0555	0.9
	2	1761	5.0	0.0559	9.4	0.0518	7.5
	3	1803	2.7	0.0636	-3.1	0.0544	2.9
	Average	1800	2.9	0.0622	-0.8	0.0539	3.8
10	1	1722	7.1	0.0667	-8.1	0.0565	-0.9
	2	1833	1.1	0.0632	-2.4	0.0552	1.4
	3	1689	8.8	0.0587	4.9	0.0567	-1.3
	Average	1748	5.7	0.0629	-1.9	0.0561	0.2
32	1	1656	10.6	0.0615	0.3	0.0536	4.3
	2	1596	13.9	0.0629	-1.9	0.0567	-1.3
	3	1644	11.3	0.0616	0.2	0.0581	-3.8
	Average	1632	11.9	0.0620	-0.5	0.0561	-0.2
100	1	1644	11.3	0.0652	-5.7	0.0536	4.3
	2	1626	12.2	0.0654	-6.0	0.0566	-1.1
	3	1554	16.1	0.0601	2.6	0.0550	1.8
	Average	1608	13.2	0.0636	-3.1	0.0551	1.6
320	1	1464	21.0	0.0702	-13.8	0.0570	-1.8
	2	1614	12.9	0.0642	-4.1	0.0567	-1.3
	3	1482	20.0	0.0755	-22.4	0.0601	-7.3
	Average	1520	18.0	0.0700	-13.5	0.0579	-3.4
1000	1	1485	19.9	0.0675	-9.4	0.0584	-4.3
	2	1533	17.3	0.0751	-21.7	0.0592	-5.7
	3	1584	14.5	0.0751	-21.7	0.0604	-7.9
	Average	1534	17.2	0.0725	-17.5	0.0593	-5.9

 

Calculated EC50 and NOEC

Based on I_Avalue

	(mg/L)	95-Percent Confidence Limits (mg/L)
EbC50 (0-72h)	>1000	-
NOECb (0-72h)	10	-

Based on I_mvalue

	(mg/L)	95-Percent Confidence Limits (mg/L)
ErC50 (24-48h)	>1000	-
NOECr (24-48h)	1000	-
ErC50 (24-72h)	>1000	-
NOECr (24-72h)	1000	-

 

Temperature In Algal Culture Cabinet

Temperature (°C)
0h	24h	48h	72h	Average
23.1	23.1	23.1	23.1	23.1

 

pH Values at 0-Hour and 72-Hour Exposure

Nominal Concentration (mg/L)		pH
		0 Hour	72 Hour
Control	1 2 3	8.6	9.4 9.8 9.6
1.0	1 2 3	8.6	9.7 10.1 10.0
3.2	1 2 3	8.3	9.8 9.6 9.9
10	1 2 3	8.4	9.5 9.8 9.6
32	1 2 3	8.4	9.8 9.9 9.7
100	1 2 3	8.5	9.3 9.7 9.6
320	1 2 3	8.3	9.4 9.1 9.6
1000	1 2 3	8.3	9.7 9.3 9.5

Validity criteria fulfilled:

yes

Conclusions:

ErC50 (24- 72h): > 1000 mg/L
No Observed Effect Concentration (NOECr): 1000 mg/L

Executive summary:

Purpose of the study: Growth Inhibition test to algae (Selenastrum capricornutum) of sodium p-toluenesulfonate will be conducted to determine growth inhibition concentration (EC50) and No Observed Effect Concentration (NOEC).

Applied Guideline: This study is done in accordance with OECD Guidelines for the testing chemicals No.201 "Alga, Growth Inhibition test" (1984).

Applied GLP: This study is done in accordance with GLP regulation of Environment Agency.

1) Test substance: Sodium p-toluenesulfonate

2) Incubation: Shaking cultivation (100 rpm)

3) Test organism: Selenastrum capricornutum (ATCC 22662)

4) Temperature: 23±2°C

5) Test duration: 72 hours

6) Volume of test solution: 100mL (OECD medium)

7) Lighting: 4000- 5000 lux (continuous lighting)

8) Initial cell concentration: 1 X 10⁴cells/ml

9) Test Concentration: Control, 1000, 320, 100, 32, 10, 3.2 and 1.0 mg/L

10) Analysis method of test substance in the test solution: HPLC (start and end of the exposure)

 

Results

1) Growth Inhibition Concentration based on comparison of area under growth curves.

EbC50 (0-72 h): > 1000 mg/L

No Observed Effect Concentration (NOECb): 10mg/L

2) Growth Inhibition Concentration based on comparison of growth rate.

ErC50 (24-48h): > 1000 mg/L

No Observed Effect Concentration (NOECr): 1000 mg/L

ErC50(24- 72h): > 1000 mg/L

No Observed Effect Concentration (NOECr): 1000mg/L

Description of key information

The growth inhibition of algae was determined acording to the OECD 201 test guideline in accordance with GLP, with exposure to Selenastrum capricornutum under static conditions at a test concentration of 1000 mg/l.  As the substance is highly water soluble and hydrolytically stable no analytical monitoring was conducted.  Following a 72 hour exposure, no immobilisation occured.  The ErC50 is >1000 mg/l.

Key value for chemical safety assessment

EC50 for freshwater algae:

1 000 mg/L

EC10 or NOEC for freshwater algae:

1 000 mg/L