Diammonium tartrate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Please refer to the analogue justification provided in IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 33 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Source: Ref. 6.1.3-1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Source: Ref. 6.1.3-2

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 - 12 Jan 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations and sampling method: 10 mL samples were collected at timepoints 0, 24 and 48 h from both the freshly prepared and the old media for chemical analysis. Old test media were pooled prior to sampling. Additionally a sample from the 100 mg/L test concentration prepared without organisms was taken for analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 500 mg of test substance was added to 1000 mL of medium. The solution was filtered through a 0.45 μm filter. A sample of the filtrate was taken for chemical analysis. Based on the measured concentration of the filtrate, a dilution was prepared in medium resulting in the 100 mg/L test concentration. From this concentration a serial dilutions was performed in order to prepare the remaining test concentrations.
- Evidence of undissolved material and development of method for preparation of test solutions: prior to the test beginning it was observed that the test substance formed a dispersion in the test medium. Therefore the test medium with test substance was filtered. However, the test substance analysis showed that no test substance was present in the filtered sample. Therefore a solubility test and a medium preparation trial was performed:
- SOLUBILITY TEST: nominal concentrations of 100, 500 and 1000 mg/L were prepared and filtered through a 0.45 μm filter. Duplicates were chemicaly analysed.
- MEDIA PREPARATION TRIAL: A 500 mg/L stock solution was prepared, in duplicate, and filtered through a 0.45 μm filter. Duplicate samples were chemicaly analysed. Analysis was repeated after 24 and 48 h. Based on these results, the test solutions for the actual test were prepared as described under `method´.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Age of parental stock: < 24 h old
- Feeding during test: no

ACCLIMATION
- Acclimation conditions: 24 h before a test was set up, juveniles present in the cultures were removed. Over the next 24 h, juveniles for use in the test were removed from the culture and transferred to fresh culture medium. Jjuveniles were then left for at least 1 h. Actively swimming individuals were chosen for the test.

HOLDING
- Type and amount of food: concentrated suspension of Chlorella vulgaris prepared in accordance with standard operating procedures
- Feeding frequency: daily

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

-

Test temperature:

18.6 - 19.8

pH:

7.35 - 7.69

Dissolved oxygen:

87 - 98% of air saturation value
8.39 - 9.58 mg/L

Salinity:

-

Conductivity:

-

Nominal and measured concentrations:

Nominal: 0 (control), 10, 18, 32, 56 and 100 mg/L
Measured: < LOQ, 6.5, 9.3, 11, 15 and 33 mg/L (time-weighted mean measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass tall form beakers covered with a lid
- Fill volume: 50 mL
- Renewal rate of test solution: 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per abiotic control (replicates): 1 vessel for the 100 mg/L test item concentration


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH, dissolved oxygen concentration and temperature were determined in freshly prepared test media and in the old media at 24 and 48 h.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16-hour light: 8-hor dark (fluorescent lighting)

EFFECT PARAMETERS MEASURED: mobility was observed after 24 and 48 h

RANGE-FINDING STUDY
- Test concentrations: 1, 10 and 100 mg/L (nominal)
- Results used to determine the conditions for the definitive study: no short term toxicity was observed with an EC50 (48 h) of > 100 mg/L (nominal)

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 33 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: no mortalities
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: although the initial 500 mg/L stock solution was a colourless solution, the 10 mg/L test concentration was observed to be very slightly cloudy, the 18-56 mg/L test concentrations were slightly cloudy and the 100 mg/L test concentration was cloudy, non-homogenous dispersions with some precipitates. This was contrary to the observations from the range-finding/limit test and the media preparation test where prepared concentrations were observed to be colourless solutions. It was considered possible that the test substance was reacting with the salts dissolved in the test media. This was considered not to affect the validity of the test given that no immobilisation was observed during the test which was consistent with the range finding test. The control was a colourless solution throughout the test.

SOLUBILITY TEST

Table 1: Results from the solubility test.

Initial concentration (mg/L)	Sample number	Measured concentration (mg/L)	Mean measured concentration (mg/L)
100	1	<LOQ	<LOQ
	2	<LOQ
500	1	280	258
	2	236
1000	1	329	338
	2	347

LOQ – Limit of quantification assessed as 5 mg/L

Based on the results of the solubility test, it was considered appropriate to prepare the test solutions from an initial 500 mg/L stock solution followed by filtration through a 0.45 μm filter. The resulted filtrate was analysed to determine the concentration prior to dilution and give the highest test concentration of 100 mg/L.

ANALYTICAL RESULTS

Table 2: Test item concentrations in the media during the test.

Nominal concentration (mg/L)	0 h (New media)	% nominal	24 h (Old media)	% nominal	24 h (New media)	% nominal	48 h (Old media)	% nominal	Time weighted mean measured concentration (mg/L)**
Control	<LOQ	-	<LOQ	-	<LOQ	-	<LOQ	-	-
10	10.25	103	5.95	60	8.93	89	<LOQ	-	6.5
18	17.94	100	5.13	29	20.13	112	<LOQ	-	9.3
32	26.55	83	<LOQ	-	36.31	113	<LOQ	-	11
56	43.3	77	<LOQ	-	51.52	92	<LOQ	-	15
100	82.28	82	8.46	8	104.89	105	5.35	5	33
100*	NA	NA	10.18	10	NA	NA	8.62	7	NA

* Vessel without test organisms

** Where measured concentrations were <LOQ, a value of half LOQ (2.5 mg/L) was used to calculate the time-weighted mean measured concentration

The 100 mg/L test concentration without test organisms, gave similar results to the samples from media containing test organisms. This indicated that the decline of the test substance in the medium during the test period was possibly due to chemical and not biological reasons.

Due to the decrease of the measured concentrations over a 24-hour period, it was considered appropriate to base the results on time-weighted mean measured test concentrations.

BIOLOGICAL DATA

Table 3: Numbers of immobilised Daphnia magna after 48 h exposure.

Nominal concentration (mg/L)	Time-weighted mean measured concentration (mg/L)	% Immobility at 48 h
Control	Control	0
10	6.5	0
18	9.3	0
32	11	0
56	15	0
100	33	0

Table 4: Further toxicity results.

	Time-weighted mean measured concentration (mg/L)
LOEC (48 h)	>33
NOEC (48 h)	≥33

Validity criteria fulfilled:

yes

Remarks:

control immobility was ≤10% and dissolved oxygen was ≥3 mg/L

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28-30 April 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

4.8 mL samples for possible analysis were taken from all test concentrations and the control at t=0 and t=48 hours and were stored in the freezer until analysis.

Vehicle:

no

Details on test solutions:

Preparation of test solutions started with the highest concentration (100 mg/L) applying 30 minutes of magnetic stirring to accelerate the dissolving of the test substance in the test medium. The lower test concentrations were prepared by subsequent dilutions of the stock in test medium. The final test solutions were all clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Dapnia magna
- Source: In-house laboratory culture with a known history, at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.
- Age at study initiation (mean and range, SD): young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.
- Method of breeding: start of each batch with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel. Renewal of the cultures: after 7 days of cultivation half of the medium twice a week.
- Feeding during test: no

ACCLIMATION
- Acclimation period: <24 hours
- Acclimation conditions (same as test or not): no, breeding medium is M7
- Type and amount of food: a suspension of fresh water algae
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

None

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

18.7 - 19.4 ºC

pH:

7.7-8.0

Dissolved oxygen:

8.6-9.1 mg/L

Nominal and measured concentrations:

Nominal: 0.10, 1.0, 10 and 100 mg/L
At 100 mg/L, the actual concentration was at the level of nominal (109%) at the start of the test and remained stable during the test (99% of initial). Consequently, the effects parameters were reported in terms of the analytically confirmed nominal concentration.

Details on test conditions:

TEST SYSTEM
- Test vessel:100 mL, all-glass, fill volume 80 mL
- Aeration: None
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for the limit concentration, 2 for the remaining concentrations
- No. of vessels per control (replicates): 4
- Biomass loading rate: 5 per vessel containing 80 mL test solutionn

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium
- Culture medium different from test medium: Culture medium had extra trace elements, macro nutrients and vitamins added as specified in Medium M7.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours per day, During the first 24 hours of the study the lights were not turned off during the night. Therefore, the light period was longer than 16 hours during the first 24 hours.
- Light intensity: not indicated

EFFECT PARAMETERS MEASURED: mobility at 24 and 48 hours ; pH and dissolved oxygen at the beginning and at the end of the test. Temperature continuously.

TEST CONCENTRATIONS
- Test concentrations: combined limit/range-finding test: 0.10, 1.0, 10 and 100 mg/L.

Reference substance (positive control):

yes

Remarks:

Potassiumdichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Nominal test concentrations were analytically confirmed.

Details on results:

No effects were observed in any of the groups tested.

Results with reference substance (positive control):

The 48h-EC50 was 0.32 mg/L with a 95% confidence interval between 0.29 and 0.37 mg/L.

Validity criteria fulfilled:

yes

Remarks:

1. No effects were observed in the control. 2. The oxygen concentration at the end of the test was ≥3 mg/l in control and test vessels

Conclusions:

No effects were observed in any of the groups. The 48h-EC50 of strontium tartrate to Daphnia magna is > 100 mg/L.

Executive summary:

A 48-h acute toxicity study in Daphnia magna was performed with Strontium tartrate according to OECD 202 and in compliance with GLP principles. A combined range-finder/ limit test was carried out under static conditions. The nominal test concentrations wer 0.10, 1.0, 10 and 100 mg/L. Preparation of test solutions started with the highest concentration (100 mg/L) applying 30 minutes of magnetic stirring to accelerate the dissolving of the test substance in the test medium. The lower test concentrations were prepared by subsequent dilutions of the stock in test medium. The final test solutions were all clear and colourless. Samples were taken for analysis at t = 0 and 48 hours. Samples of the control and 100 mg/L group were analysed by an ICP-MS method. At 100 mg/L, the actual concentration was at the level of nominal (109%) at the start of the test and remained stable during the test (99% of initial). Consequently, the effects parameters were reported in terms of the analytically confirmed nominal concentration.

During the first 24 hours of the study the lights were not turned off during the night. Therefore, the light period was longer than 16 hours. The results show that the daphnids were not affected (no immobility observed). Furthermore, at the end of the test the test substance concentration remained at the initial concentration (99% of initial) indicating that the light period did not influence the test substance concentration.

No effects were observed in any of the groups. The 48h-EC50 of the substance to Daphnia magna is > 100 mg/L.

Description of key information

EC50 (48 h) > 100 mg/L (nominal, OECD 202) based on read across from strontium tartrate (CAS 868-19-9)

EC50 (48 h) > 33 mg/L (meas. TWA, OECD 202) based on read across from diammonium oxalate (CAS 6009-70-7)

Additional information

Experimental data on the toxicity of diammonium tartrate (CAS 3164-29-2) on aquatic invertebrates were not available. The toxicity was assessed using read across data from the analogues substances strontium tartrate (CAS 868-19-9) and diammonium oxalate (CAS 6009-70-7). Details on the read across justification are provided in section 13 of the IUCLID dossier.

The study with the read across substance strontium tartrate (reference: 6.1.3-2) was conducted according to the OECD guidance 202 and GLP standards. In a static system the test organism Daphnia magna was exposed for 48 h to the nominal test item concentrations of 0 (control), 0.10, 1.0, 10 and 100 mg/L. The immobility of the water flea was determined after 24 and 48 h. The test item concentrations were analytically verified using ICP-MS at the beginning and at the end of the test. The analytical recovery of the test substance in the medium was between 99 and 109%. Therefore the results were expressed in nominal concentrations. No toxic effects were observed up to the highest tested concentration resulting in an EC50 (48 h) of > 100 mg/L (nominal). On the basis of these results it is concluded that neither the tartrate anion nor the metal strontium, reveal toxic effects to Daphnia up to the highest tested concentration. Regarding the toxicity of the non-common compound - strontium -between the target and the source substance, findings by Okamoto et al. 2014, who tested the toxicity of 50 metal compounds to Daphnia magna according to OECD guideline 202, also concluded low acute toxic effects by strontium starting at concentrations higher than 120 mg pure strontium per L.

Potential toxic effect by the ammonium species attached to the tartrate, is evaluated based on experimental data of the analogues substance diammonium oxalate in a read across approach. The study was conducted according to the OECD guidance 202 and GLP standards (reference: 6.1.3-1). In a semi-static system the test organism Daphnia magna was exposed for 48 h to nominal test item concentrations of 0 (control), 10, 18, 32, 56 and 100 mg/L (corresponding to the measured concentrations of < LOQ (control), 6.5, 9.3, 11, 15 and 33 mg/L, expressed in TWA). Chemical analysis of the test media samples was conducted at 0 (fresh media), 24 (old and fresh media) and 48 h (old media). Since the recovery of the test substance was between < LOQ and 60% after 24 h incubation, the test results were expressed in means of the time weighted average. No toxic effects to Daphnia magna were observed up to the highest tested concentration resulting in an EC50 (48 h) of > 33 mg/L (meas. TWA).

The extensive report by EPA concerning the aquatic life ambient water quality criteria for ammonia, (2013), summarizes literature data among others on the toxicity of ammonium nitrogen to aquatic invertebrates. All collected toxicity values are normalized to pH 7.0 and temperature of 20°C. 52 species of invertebrates were used for toxicity assessment. The effect values were in the range between 23.12 (Venustaconcha ellipsiformis) and 2515 (Erythromma najas) mg total ammonium nitrogen/L. The final acute value (FAV), which was determined by regression analysis using a log-triangular fit based on the four most sensitive genera in the data set, was 33.52 mg total ammonium nitrogen/L. This represents a very conservative toxicity effect value for ammonium nitrogen. Extrapolating this value to diammonium tartrate, results in the very conservative effect value corresponding to 220.34 mg diammonium tartrate/L, while effects are attributed in this case to the ammonium concentrations. This calculation is in line with the results obtained from the study with the read across substance diammonium oxalate (no toxicity up to the highest tested concentration of 100 mg diammonium oxalate/L) and shows that no concern from ammonium toxicity arises up to 100 mg diammonium tartrate/L for aquatic invertebrates.

In conclusion based on the results of the read across substances and the literature data, no adverse effects are expected by diammonium tartrate towards aquatic invertebrates.

Reference:

Okamoto A., Yamamuro M., Tatarazako N. 2014 Acute toxicity of 50 metals to Daphnia magna: Acute toxicity of 50 metals to D. magna.Journal of Applied Toxicology. 35: 824-830

United States Environmental Protection Agency. 2013. Aquatic life ambient water quality criteria for ammonia – freshwater. EPA-822- R-13-001. Office of Water, Office of Science and Technology, Washington, DC.