A mixture of: N,N-di(hydrogenated alkyl C14-C18)phthalamic acid; dihydrogenated alkyl (C14-C18)amine

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:

1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

87787-81-3

EC Number:

618-070-1

Cas Number:

87787-81-3

IUPAC Name:

87787-81-3

Constituent 2

Reference substance name:

A mixture of: N,N-di(hydrogenated alkyl C14-C18)phthalamic acid; dihydrogenated alkyl (C14-C18)amine

EC Number:

413-800-3

EC Name:

A mixture of: N,N-di(hydrogenated alkyl C14-C18)phthalamic acid; dihydrogenated alkyl (C14-C18)amine

Cas Number:

87787-81-3

Molecular formula:

C44H79NO3

IUPAC Name:

2-(dioctadecylcarbamoyl)benzoic acid; 2-(ditetradecylcarbamoyl)benzoic acid; 2-[(propan-2-yloxy)carbonyl]benzoic acid; 2-[octadecyl(tetradecyl)carbamoyl]benzoic acid; 2-octadecyl-2,3-dihydro-1H-isoindole-1,3-dione; 2-tetradecyl-2,3-dihydro-1H-isoindole-1,3-dione; benzene-1,2-dicarboxylic acid; dioctadecylamine; ditetradecylamine; octadecyl(tetradecyl)amine

Constituent 3

Reference substance name:

reaction mass of: N,N-di(hydrogenated alkyl C14-C18)phthalamic acid dihydrogenated alkyl (C14-C18)amine

IUPAC Name:

reaction mass of: N,N-di(hydrogenated alkyl C14-C18)phthalamic acid dihydrogenated alkyl (C14-C18)amine

Constituent 4

Reference substance name:

Stepan TAB-2

IUPAC Name:

Stepan TAB-2

Details on test material:

Test substance was treated as 100% pure. Test substance is stable at 50 deg C and Stored at room temperature in the dark. Expiry date April 30, 1992. Test substance batch number: 198308. Test substance is stable for at least 48 hours in water.

Specific details on test material used for the study:

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

Sampling and analysis

Analytical monitoring:

not specified

Details on sampling:

No detail other than what was described under test conditions.

Test solutions

Vehicle:

yes

Details on test solutions:

Composition of medium M4:
ISO-medium: The following chemicals (analytical grade) are dissolved in freshly prepared ultra-pure water1) (mg/l):
CaCl2 .2H2O 293,8
MgSO4.7 H2O 123.3
NaHCO3 64.8
KCl 5.8
Medium M4: Trace elements, macro nutrients and vitamins are added to freshly prepared ISO-medium to reach the following concentrations:

Trace elements (mg/l)
B 0.5
Fe 0.2
Mn 0.1
Li, Rb and Sr 0.05
Mo 0.025
Br 0.0125
Cu 0.0063
Co and I 0.0025
Se 0.0010
V 0.0003
Na2EDTA.2H2O 2.5

Macro nutrients (mg/l
Na2SiO3.9H2O 10
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18

Vitamins (µg/l)
Thiamine 75.0
B12 1.0
Biotin 0.75
1)Tap water purified by reverse osmosis and subsequently passed over activated carbon and ion-exchange cartridges: Milli-Q water (Millipore Corp., Bedford, Mass., USA).
Temperature of Medium: 18-22, constant within ±1°C.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia were of an age of <24 hours. The start of each batch was with newborn animals, i.e., less than 3 days old, by placing about 250 of them into 10 liters of medium in an all glass culture vessel. The maximum age of the cultures was 4 weeks. Renewal of the cultures: After 7 days of cultivation half of the medium was renewed twice a week. Feeding was daily, with a suspension of fresh water algae.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

No remarks.

Post exposure observation period:

No data provided in study report.

Test conditions

Hardness:

No data provided in study report.

Test temperature:

Temperature of Medium: 18-22°C, constant within ±1°C. During the study the temperature of the test medium measured in the blank control decreased from 20 to 18.5°C.

pH:

At the start of the final test pH decreased from 8.0 to 7.0 with increasing concentration. After 48 hours pH ranged from 8.3 to 8.1 with increasing concentration.

Dissolved oxygen:

Oxygen concentration in the test media was found to be >5 mg/l for all measurements.

Salinity:

No data provided in study report.

Nominal and measured concentrations:

Based on the results of the pre-test daphnia were exposed to nominal concentrations of 100, 180, 320, 560, and 1000 mg/l.

Details on test conditions:

Frequent inspections were conducted of the cultures with respect to number of young, appearance of young and parental daphnia and global feeding behavior.

The test was combined with a reference test with potassium dichromate in DSW and one in M4.

Initially a pre-test was performed in which exposure was limited to the maximum concentration of nominally 100 mg/l. Hence daphnia were exposed for 48 hours to a supersaturated solution of 1000 mg/l and its filtrate. The test was performed in duplicate with 10 daphnia per vessel.
Test type: static.
Test vessels: 250 ml, all glass
Medium: M4
Number of daphnia: 20 per concentration.
Light: 16 hours photoperiod daily
Feeding: no feeding during exposure period

Controls: Test medium without test substance or other additives (0 mg/l) and one control containing test medium with the additive used in the treatment of the stock solutions, i.e., Tween 80 (0.1 ml/l).

Introduction of young daphnia: The daphnia were introduced into the test medium directly after preparation of the test media.
The test media were prepared in M4-medium. Because of the low solubility (<1 mg/L), a mixture of the test substance (heated to 50°C before use) and Tween 80 was added quantitatively to M4 (50°C) providing concentrations of nominally 100, 180, 329, 560 and 1000 mg. These solutions were then cooled to about 20°C. In all test solutions oily-like drops were floating on the surface.

Measurements and Recordings:
Immobility (including mortality): At 24 hours (except in the solutions containing 1000 mg/l due to turbidity and test substance floating on the surface of the medium) and at 48 hours. At 48 hours the daphnia present in the solution containing 1000 mg/l were checked individually for mobility by taking them out of the solution with a pipette.
pH and Oxygen: Measured at the beginning and at the end of the test for all concentrations and the controls

Temperature of Medium: Determined daily in one control vessel, beginning at the start of the test.

Acceptability criterion of the test: In the control, or more than 10% of the daphnia should have been immobilized or trapped at the surface of the water. This criterion was met.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 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:

Pre-Test: In the pre-test 40% immobilization was recorded in the supersaturated solution and 30% immobilization in its filtrate.

Final Test: After 24 hours of exposure no immobilization of daphnia was seen at any of the concentrations tested up to and including 560 mg/l. At 1000 mg/l no possible effect on mobility could be recorded due to the opacity of the test solution. After 48 hours of exposure 10% immobilization was seen at 1000 mg/l while no immobilization of daphnia was seen at or below 560 mg/l.

In the controls no daphnia became immobilized or trapped at the surface of the water.

Under the conditions of the final test, the 48 hour EC50 for immobility of daphnia exposed to Stepan TAB-2 was higher than the nominal 1000 mg/l, at which concentration no complete mixture in water could be achieved. Finally considering a total effect of 10% as not biologically significant, the No Observed Effect Concentration was nominally 1000 mg/l.

Results with reference substance (positive control):

Reference tests: (RCC NOTOX PCDA-E56): 48-hour acute toxicity study in the daphnia with potassium dichromate. This reference test was carried out in 1991 to check the sensitivity of the test system as used by RCC NOTOX. The reference substance potassium dichromate was tested under similar conditions at the same time as the final test with Stepan TAB-2 using daphnia from the same culture. The concentrations were 0.10, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/l in M4 medium. The control was Medium M4 without test substance. The 24 hour EC50 was between 1.0 and 1.8 mg/l. The 48 hour EC50 was between 0.56 and 1.0 mg/l. A reference test with potassium dichromate in Dutch Standard Water was also performed simultaneously with the following results: The 24 hour EC50 was 0.95 mg/l, which approximates the lower limit of the expected range: 1.0-2.4 mg/l (n=60). The 48 hour EC50 was between 0.56 and 1.0 mg/l, which is within the expected range: 0.6-1.9 mg/l (n=60). Hence the sensitivity of the daphnia for the reference substance was as expected rom the historical data.

Reported statistics and error estimates:

No data provided in the study report.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Validity of the test: In the control, or more than 10% of the daphnia should have been immobilized or trapped at the surface of the water. This criterion was fulfilled.

Conclusions:

Under the conditions of the short-term daphnia study, the 48-hour EC50 if Stepan TAB-2 was determined to be >1000 mg/l, at which concentration no complete solution in water could be achieved.

Executive summary:

The water solubility of Stepan TAB-2, using the column elution method, was determined to be <0.1 mg/l (RCC NOTOX Project 058883)(Summarized elsewhere in this substance file).

Daphnia were exposed for a maximum of 48 hours to a concentration range of 100 -1000 mg/l forming a geometric progression with a factor of 1.8. Because of the low solubility of the test substance, Tween 80 was used as an emulsifier (0.1 ml/l)/ The test was performed in duplicate with 10 daphnia per vessel.

After 24 hours no immobilization of daphnia was seen at any of the concentrations up to and including 560 mg/l. At 1000 mg/l no possible effect on mobility could be recorded due to the opacity of the test solution.

After 48 hours of exposure 10% immobilization was seen at 1000 mg/l, while no immobilization of daphnia was seen at or below 560 mg/l.

Hence, the 48 hour EC50 for immobility of daphnia exposed to Stepan TAB-2 was higher than nominally 1000 mg/l, at which concentration no complete mixture in water could be achieved.

Finally, considering a total effect of 10% as not biologically significant, the no observed effect concentration (NOEC) was nominally 1000 mg/l.