1,3,5-triazine-2,4,6-triamine phosphate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2014-07-22 to 2014-12-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Batch identification: 0010518011
- Expiry date: 17 Sep 2019
- Storage conditions: The test substance was stored at ambient temperature.

Analytical monitoring:

yes

Details on sampling:

The concentration of the test substance was determined in one representative test interval per week in each freshly prepared test solution (test vessels without daphnids) and in the 48 h or 72 h old test solutions before renewal (combined sample from test vessels with
daphnids).
Additionally, samples were collected from the 48 hour or 72 hour old abiotic replicate (no
Daphnia or food algae) per test group.

Vehicle:

no

Details on test solutions:

All test solutions were prepared individually (differential loading). The test substance was weighed into a 1 L, 2 L or 5 L volumetric flask and filled to the mark with test medium. The solution was then stirred for approximately 1 O min. The test stock solutions were visibly colorless and clear after preparation.

Test organisms (species):

Daphnia magna

Details on test organisms:

- Test species: Daphnia magna STRAUS
- Reason for selection of the test species: Recommended species in the test guidelines
- Origin : The clone of Daphnia magna STRAUS 1820 used was supplied by the lnstitut National de Recherche Chimique Appliquee, France, in 1978. From this date on this clone was cultured and bred continuously in the Ecotoxicology Laboratory of Experimental Toxicology and Ecology, BASF SE, Ludwigshafen Germany.
- Culture conditions: Daphnia brood stock are kept in mass cultures consisting of approx. 20 - 30 individuals for a maximum of 4 weeks. All individuals in the mass culture originate from a single female. After approximately 14 days the adults have produced at least 3 broods and the young can be used in tests. Offspring are removed from the mass cultures at least once daily during the normal work weelk to ensure that young daphnia are <24-h old (first instar) at test initiation. Detailed records are kept (in test facility archives) to monitor the health of Daphnia brood stock cultures including observations of young production, mortality, ephippia, and measurement of water ch,emistry parameters. Only young from healthy cultures without signs of stress are used for testing.
- Acclimatization: The Daphnia are cultured under the identical conditions as the test including test media (Elendt M4 ), water quality, temperature (20 ±1 °C), and diet.
- Age at test initiation: < 24 h
- Reference substance testing: In order to verify that the Daphnia magna culture is responding normally to toxic stress, tests with a reference substance are conducted monthly. Reference substance tests are conducted generally according to the OECD 202 guideline and in accordance with GLP, but without a GLP status. The EC50(48 h) of the reference substance sodium chloride (NaCl) was 5.25 g/L. This result is wiithin the range of 3.88 - 7.22 g/L, which represents ±2 standard deviations from the published ECso(48 h) of 5.55 g/L and indicates that the culture of Daphnia magna used in this study is responding normally to toxic stress.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

22 d

Remarks on exposure duration:

None

Post exposure observation period:

Not performed.

Hardness:

2.20 - 3.20 mmol/L

Test temperature:

20-21 deg C

pH:

7.4-8.2

Dissolved oxygen:

8.3-9.1 mg/L

Salinity:

n/a

Conductivity:

550 - 650 μS/cm

Nominal and measured concentrations:

0, 0.46, 1.0, 2.2, 4.6, 10 mg/L as nominal concentrations
0, 0.44, 0.98, 2.20, 4.83, 10.41 mg/L as time-weighted mean measured

Details on test conditions:

Test medium:
A synthetic fresh water (Elendt M4) is used as media for culture and test purposes. The general properties of this medium are as follows.
Total hardness: 2.20 - 3.20 mmol/L
Acid capacity up to pH 4.3: 0.80 -1 .00 mmol/L
Molar ratio Ca:Mg: about 4 : 1
pH value: 7.5 - 8.5
Conductivity: 550 - 650 μS/cm
Total organic carbon: < 2 mg/l
Dissolved oxygen: Must remain ~ 3mg/L during the test. To
assure optimal dissolved oxygen levels, the M4 medium is aerated for approximately 24 h prior to use.

Test vessels : Numbered glass beakers (nominal volume 100 mL), covered with glass Petri plates to slow evaporation.
Test volume: 50 mL
Biological loading: 1 animal / test vessel (0.02 animals / mL)
Light intensity / Photo period: 704 - 728 lux at a wave length of 400 -750 nm. The light intensity will not exceed 1000 - 1500 lux for cool white light (equivalent to 15-20 μE·m·2·s·1 according to the test guideline. 16 hours light : 8 hours darkness
Aeration: none
Test solution renewal: Semi-static, renewal 3 times per week
Diet:
During the test daphnids were fed daily a diet of live green algae Desmodesmus subspicatus, cultured in a synthetic
medium. The algae were separated from their culture medium by centrifugation, resuspended in daphnid's medium (M4)
corresponding to concentrations of 1760 mg and 2160 mg TOC*/L (respectively) in the algal concentrates used. The
daphnids were fed a defined volume (s114 μl) of the concentrate to reach the amount of food. The algae were stored in a refrigerator (dark, about 4- 80 °C) for maximum 21 days.
By adding the algal concentrate the test solution was slightly diluted. During a 72 h test interval with 3 feedings, 0.114 mL each, a total of 0.342 ml are added to 50 ml test volume resulting in a maximal dilution of 0.7 %.

Reference substance (positive control):

yes

Remarks:

sodium chloride

Key result

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other:

Remarks:

Since the measured concentrations of the test substance in the test solutions were within ±20% of the nominal concentrations, the effect concentration can be expressed relative to the nominal concentration.

Key result

Duration:

22 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other:

Remarks:

Since the measured concentrations of the test substance in the test solutions were within ±20% of the nominal concentrations, the effect concentration can be expressed relative to the nominal concentration.

Details on results:

There was no significant adverse effect on reproduction in any treatment group after 22 days. The data were not sufficient to calculate ECx values for reproduction. No toxic effects could be observed in a 22d reproduction test with Daphnia magna. The validity criteria were fulfilled.

No mortality was observed among parent animals over the 22d exposure period.

No additional adverse effects or abnormal behavior were obseNed in any of the test groups.

The validity criteria were fulfilled.

Results with reference substance (positive control):

The EC50(48 h) of the reference substance sodium chloride (NaCl) was 5.25 g/L. This result is within the range of 3.88 – 7.22 g/L, which represents ±2 standard deviations from the published EC50 (48 h) of 5.55 g/L and indicates that the culture of Daphnia magna used in this study is responding normally to
toxic stress.

Description of key information

The chronic toxicity to aquatic invertebrates was assessed in a study (BASF SE, 2014) conducted according to OECD 211 (2012). Daphnia magna was exposed to the test substance for 22 days. The NOEC based on reproduction was determined to be > 10 mg/L. The chronic toxicity and the reproduction toxicity of melamine to Daphnia magna is low.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Effect concentration:

> 10 mg/L

Additional information

A study with the target substance was available (BASF SE, 2014). The chronic toxicity to aquatic invertebrates was assessed in a study conducted according to OECD 211 (2012). Daphnia magna was exposed to nominal concentrations of 0, 0.46, 1.0, 2.2, 4.6, 10 mg/L test substance for 22 d. The test was conducted as semi-static test. After the exposure time no effects on any of the observed endpoints could be detected. The NOEC based on reproduction was determined to be > 10 mg/L and the LOEC >= 10 mg/L. No significant mortality or any other additional significant adverse effects or abnormal behavior were observed in any of the test treatments. The data were not sufficient to calculate ECx values for reproduction or mortality.

 

In addition, a weight of evidence approach with the read across source substance melamine (CAS 108-78-1) is applied (please refer section 13 for the read across justification of the test substance). In the supporting study (Adema, 1978 (DSM) performed with the read across substance melamine (CAS 108 -78 -1) a 21 d-NOEC for reproduction and mortality was found to be 18 mg/L.

As the substance is a salt of phosphate, effects were read-across from the cation, which is likely to have the more important effects. In biological systems, phosphate is an abundant in cells and body fluids. In ecological terms, because of its important role in biological systems, phosphate is a highly sought-after resource. Thus, read-across from component CAS 108-78-1, melamine, is justified. All effect values are minimum values as the molar ratio anion:cation equals 1.74