Asphalt, sulfonated, sodium salt

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-07-07 to 2004-07-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study performed according to Guideline United Kingdom Proposal to ISO TC147/SC5/WG2 "Water Quality, Determination of Acute Lethal Toxicity to Marine Copepods (Copepoda, Crustacea)".

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: The method used followed the recommendations of the United Kingdom Proposal to ISO TC147/SC5/WG2 "Water Quality, Determination of Acute Lethal Toxicity to Marine Copepods (Copepoda, Crustacea)"

Deviations:

yes

Remarks:

During holding, the test organisms were fed a mixture of different marine algae other than those specified in the test method. This deviation is judged not to affect the results of the study.

Principles of method if other than guideline:

N/A

GLP compliance:

yes (incl. QA statement)

Remarks:

Department of Health of the Government of the United Kingdom

Test material

Test material information

Constituent 1

Reference substance name:

Asphalt, sulfonated, sodium salt

EC Number:

269-212-0

EC Name:

Asphalt, sulfonated, sodium salt

Cas Number:

68201-32-1

Molecular formula:

UVCB

IUPAC Name:

Asphalt, sulfonated, sodium salt

Details on test material:

- Name of test material (as cited in study report): Na+ sulfonated asphalt
- Molecular formula (if other than submission substance): N/A
- Molecular weight (if other than submission substance): N/A
- Smiles notation (if other than submission substance): N/A
- InChl (if other than submission substance): N/A
- Structural formula attached as image file (if other than submission substance): N/A
- Substance type: active
- Physical state: black powder
- Analytical purity: N/A
- Impurities (identity and concentrations): N/A
- Composition of test material, percentage of components: N/A
- Isomers composition: N/A
- Purity test date: N/A
- Lot/batch No.: N/A
- Stability under test conditions: N/A
- Storage condition of test material: N/A
- Other: Test material supplied by Sponsor.

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

- The test material concentrations in test preparations were not determined by analysis.

Test solutions

Vehicle:

no

Details on test solutions:

- The test water was prepared using a commercially available formulation (Tropic Marin® Synthetic Sea Salt).
- In the definitive test, 731 g was dispersed in a final volume of 20 L of water giving a nominal salinity of 31 parts per thousand, and the pH was adjusted to 8.0 using 10 M HCl.
- Test solutions were prepared by directly adding 1000 mg of test material to the synthetic seawater, which was dispersed by ultrasonication for 30 minutes and the volume adjusted to 1 liter to give the 1000 mg/L test concentration. Aliquots of the 1000 mg/L test concentration were used to prepare the remaining test concentrations: 10, 18, 32, 56, 100, 180, 320 and 560 mg/L. Each prepared concentration was inverted several times to ensure adequate mixing and homogeneity.

Test organisms

Test organisms (species):

other: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Common name: Marine copepod
- Strain: N/A
- Source: Guernsey Sea Farms, Vale, Guernsey, UK
- Age at study initiation (mean and range, SD): 11 - 14 days were supplied for use in the definitive test and positive control.
- Weight at study initiation (mean and range, SD): N/A
- Length at study initiation (length definition, mean, range and SD): N/A
- Valve height at study initiation, for shell deposition study (mean and range, SD): N/A
- Peripheral shell growth removed prior to test initiation: N/A
- Method of breeding: N/A
- Feeding during test
- Food type: N/A
- Amount: N/A
- Frequency: Not fed during test


ACCLIMATION
- Acclimation period: 1 day
- Acclimation conditions (same as test or not): sameas test; The test animals were held in 20 +/- 1 deg. C.
- Type and amount of food: Held in sea water with a salinity of 33 +/- 2 parts per thousand and fed on a mixed culture of marine algae. Algal mix used as food consisted of Pavlova lutheri and Tetraselmis sueccica supplied by Guernsey Sea Farms, rather than Skeletonema sp. and Rhodomonas sp. as stated in the standard test method. This deviation was considered to have no effect on the test results.
- Feeding frequency: N/A
- Health during acclimation (any mortality observed): N/A


QUARANTINE (wild caught)
- Duration: N/A
- Health/mortality: N/A

Study design

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

N/A

Test conditions

Hardness:

N/A

Test temperature:

- The water temperature was recorded daily throughout the test in each replicate test vessel.
- The test was carried out in a temperature-controlled room at 21.7 - 22.3 deg. C.

pH:

- The pH was recorded at test start and termination in each replicate test vessel.

Dissolved oxygen:

- The test vessels were not aerated during the test.
- The DO concentrations were recorded at test start and termination in each replicate test vessel.

Salinity:

- 31 ppt in dilution water

Nominal and measured concentrations:

- The test concentrations used in the definitive test were based on the results of a range-finding test.
- Based on the results of the range-finding test, the following nominal concentrations were chosen for the definitive test: 10, 18, 32, 56, 100, 180, 320, 560, and 1000 mg/L.
- The positive control was tested at: 0.56, 1.0, 1.8, 3.2, 5.6, 10, 18, 32, and 56 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL jars
- Type (delete if not applicable): closed- covered to reduce evaporation
- Material, size, headspace, fill volume: 50 mLglass jars, 25 mL test solution
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): N/A
- Renewal rate of test solution (frequency/flow rate): N/A
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): N/A
- Biomass loading rate: N/A


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic seawater made with Tropic Marin seasalt
- Total organic carbon: N/A
- Particulate matter: N/A
- Metals: N/A
- Pesticides: N/A
- Chlorine: N/A
- Alkalinity: N/A
- Ca/mg ratio: N/A
- Conductivity: N/A
- Culture medium different from test medium: N/A
- Intervals of water quality measurement: N/A


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light, 8 h dark, 20 min dawn and dusk transitions
- Light intensity: N/A


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, at 24 and 48 h. At 320, 560 and 1000 mg/L, the organisms were removed from the test vessels and placed in a Petri dish during the 24 and 48 h observations due to the turbidity of the test preparations.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Justification for using less concentrations than requested by guideline: N/A
- Range finding study
- Test concentrations: 0.1, 1.0, 10, and 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

: potassium dichromate - Exposure conditions were the same as those in the definitive test with the exception that the temperatures were shown to range from 21.7 - 22.2 deg C.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: No CL

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

380 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CL= 330 - 440 mg/L

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality

Details on results:

MORTALITY:
- At 24 h, less than 50% mortality was observed at the highest test concentration of 1000 mg/L, therefore, it was not possible to calculate an LC50 value from the data by the probit method.
- At 48 h, there was no mortality at 10 - 180 mg/L, with increasing mortality at higher concentrations.

MATERIAL SOLUBILITY:
- The turbidity of the dispersion and amount of precipitate increased with increasing concentration. The 10 mg/L treatment was a clear colorless solution throughout the duration of the test. The other treatments were observed to be brown dispersions with fine brown particles in suspension at 0 hours; these particles settled to the bottom of the vessel during the test.

OBSERVATIONS ON THE TEST ORGANISMS:
- It was confirmed that no TS adhered to the test organisms in the definitive test. Thus, mortality was not attributable to a physical effect.

PHYSICO-CHEMICAL MEASUREMENTS:
- pH ranged from 8.1 - 8.2 in all treatments throughout the test.
- Dissolved oxygen ranged from 52% to 84% of the air saturation value (ASV) at 0 hours, with decreasing ASV at higher test concentrations. At 48 hours, DO was 70 - 78% ASV at all test concentrations.
- Temperature ranged from 21.7 - 22.2 in all treatments throughout the test.

Results with reference substance (positive control):

- The 24 h LC50 was: 35 mg/L (95% CI 29 - 47 mg/L).
- The 48 h LC50 was: 4.5 mg/L (95% CI 3.8 - 5.5 mg/L).

POSITIVE CONTROL:
- The results from the positive control were within the normal range for this potassium dichromate.
- The 24 h NOEC was: 10 mg/L.
- The 48 h NOEC was: 0.1 mg/L.

PHYSICO-CHEMICAL MEASUREMENTS:
- The temperature was maintained at 21.7 - 22.3 deg C throughout the test (this is above the temperature range in the protocol; however, it was determined that there were no temperature-related effects on the test organisms).
- No treatment-related differences in pH.
- A concentration-dependent difference in oxygen concentration was observed throughout the test.

Reported statistics and error estimates:

The LC50 values and associated confidence limits at 24 and 48 hours were calculated by the maximum-likelihood probit method (Finney 1971) using the ToxCalc computer software package (ToxCalc 1991). Probit analysis is used were two or more partial responses to exposure are shown.

Any other information on results incl. tables

Table 1. Cumulative Mortality in the Definitive Test.

Cumulative Mortality Data in the Definitive Test
Nominal Concentration (mg/L)	Cumulative Mortalities Initial Population: 5 Per Replicate
	24 Hours						48 Hours
	R1	R2	R3	R4	Total	%	R1	R2	R3	R4	Total	%
Control	0	0	0	0	0	0	0	0	0	0	0	0
10	0	0	0	0	0	0	0	0	0	0	0	0
18	0	0	0	0	0	0	0	0	0	0	0	0
32	0	0	0	0	0	0	0	0	0	0	0	0
56	0	0	0	0	0	0	0	0	0	0	0	0
100	0	0	0	0	0	0	0	0	0	0	0	0
180	0	0	0	0	0	0	0	0	0	0	0	0
320	0	0	0	0	0	0	1	1	2	2	6	30
560	0	0	0	0	0	0	5	5	4	4	18	90
1000	3	2	2	1	8	40	5	5	5	6	20	100

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

sufficient survival in controls

Conclusions:

The 48 hr LC 50 of the test material to the marine copepod Acartia tonsa has been found to be 380 mg/L with 95% CL of 330 - 440 mg/L. The NOEC at 48 h was 180 mg/L.

Executive summary:

N/A