Ammonium formate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-12-07 to 2005-12-22

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:

adopted April 13th, 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

IC

Details on sampling:

- Concentrations: Nominal concentrations of the test samples: 0, 45.0, 90.0, 180, 360, 720 mg/L

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test concentrations were obtained by dilution of the highest test concentration with dilution water.
- Controls: The controls were held under the same conditions in dilution water.
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No
- Other relevant information:
Dilution water:
Reconstituted ISO test water according to the OECD-Guideline (1), annex 3, and lSO was used:
Calcium chloride solution: 11.76 g CaCl2*2H2O/1 litre with deionised water
Magnesium sulphate solution: 4.93 g MgSO4*7H2O/1 litre with deionised water
Sodium bicarbonate solution: 2.59 g NaHCO3/1 litre with deionised water
Potassium chloride solution: 0.23 g KCl / 1 litre with deionised water.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: Daphnia magna STRAUS
- Stage and instar at study initiation: 4-24h old
- Method of breeding: Adult Daphnia, at least 3 weeks old, were separated from the stock population by sieving. Batches of 30 to 50 animals were held at room temperature in approx. 1800 mL purified drinking water. During the week the daphnids were fed daily with an algal suspension (Scenedesmus subspicatus) and Liquizell® (HOBBY) according to the EEC-Guideline. Algae growing in the log-phase were centrifuged and the pellet was resuspended in a few mL of medium. 30 mL of this suspension was given to 1L Daphnia medium. The water was changed once per week. Newborn daphnids were separated by sieving, the first generation was discarded.
- Source: Umweltbundesamt, lnstitut fur Wasser-, Boden- und Lufthygiene, Berlin, Germany. Bred in the laboratory of the Fh-lME.
- Age of parental stock (mean and range, SD): at leat 3 weeks
- Feeding during test No

ACCLIMATION
- Acclimation period: at leat one week
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: algal suspension (Scenedesmus subspicatus)
- Feeding frequency: daily
- Health during acclimation (any mortality observed): No mortality reported

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

The temperature was between 20.1 and 20.3 °C

pH:

At test start, the pH in the test vessels was between 7.23 and 7.93 , after 48 h the pH was between 7.57 and 8.06.

Dissolved oxygen:

The oxygen saturation in all test concentration plots was between 90 % and 94 %

Conductivity:

10 µS x cm-1

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers
- Material, size, headspace, fill volume: 60 mL glass beakers, filled with 50 mL dilution water
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): Static
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted ISO test water according to the OECD-Guideline (1), annex 3, and lSO was used:
Calcium chloride solution: 11.76 g CaCl2*2H2O/1 litre with deionised water
Magnesium sulphate solution: 4.93 g MgSO4*7H2O/1 litre with deionised water
Sodium bicarbonate solution: 2.59 g NaHCO3/1 litre with deionised water
Potassium chloride solution: 0.23 g KCl/1 litre with deionised water.
- Culture medium different from test medium: Holding medium: Purified drinking water was used according to the OECD-Guideline. The
purification includes filtration with activated charcoal, passage through a lime-stone column and aeration. Carbonate hardness of the water is nearly 90 mg/L CaCO3, pH is in the range of 7.5 - 8.5.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16/8 light/dark
- Light intensity: the light intensity was between 713 and 736 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobility

VEHICLE CONTROL PERFORMED: no

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

360 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

207 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

365 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: Not reported
- Other biological observations: Not reported

Reported statistics and error estimates:

As the immobilization rate was 25 % at the highest test concentration an EC10 and EC50 value could not be calculated. For the assessment of the NOEC the ToxRat programme was used.
Numerical values in this report are frequently rounded to a smaller degree of precision (number of digits) than were used in the actual calculation. Minor differences in results obtained from calculations with such rounded values in comparison to those obtained with higher precision values are possible. They are however, well within the limits of the experimental accuracy and thus of no practical concern.

Concentrations of the test item at commencement and at the end of the test period.

Nominal concentration Test item [mg/L]	Test start 0 h concentration [mg/L]	Test start 0 h concentration [% nominal]	Test end 48 h concentration [mg/L]	Test end 48 h concentration [% nominal]	Geometric mean of conc. At test start and end [mg/L]	Geometric mean of conc. At test start and end [% nominal]
Control	0	-	0	-	0	-
45.0	41.0	91	39.6	88	40.3	90
90.0	80.9	90	78.4	87	79.7	89
180	163	90	140	78	151	84
360	321	89	319	88	320	89
720	636	88	629	87	633	88

Oxygen saturation and pH throughout the test

Nominal test item concentration (mg/L)	oxygen saturation				pH
	0 h		48 h		0 h	48 h
	mg/L	%	mg/L	%
Control	8.2	92	8.3	94	7.93	8.06
45.0	8.1	91	8.2	93	7.83	8.03
90.0	8.1	91	8.1	92	7.78	8.00
180	8.0	90	8.2	93	7.68	7.96
360	8.0	90	8.3	94	7.55	7.81
720	8.0	90	8.1	92	7.23	7.57

Cumulative immobility during the test period of 48 h (5 daphnids per vessel = 20 daphnids per test concentration)

Nominal concentration Test item (mg/L)	24 h				48 h				Sum total	Sum (%)
	Beaker 1	Beaker 2	Beaker 3	Beaker 4	Beaker 1	Beaker 2	Beaker 3	Beaker 4
Control	0	0	0	0	0	0	0	0	0	0
45.0	0	0	0	0	0	0	0	0	0	0
90.0	0	0	0	0	0	0	0	0	0	0
180	0	0	0	0	0	0	1	1	2	10
360	0	0	1	1	1	1	2	3	7	35
720	3	3	2	2	5	5	5	5	20	100

Validity criteria fulfilled:

yes

Conclusions:

The effect (acute immobilization) of the test item on Daphnia magna was tested using five concentrations spaced by a factor of 2. The nominal test item concentrations were: 45.0, 90.0, 180, 360 and 720 mg test item/L.
The cumulative immobility of daphnids during the test period of 48 h indicates a concentration-response relationship, ranging from 0 % immobilization at the two lowest concentrations of 45.0 and 90.0 mg test item/L to 100 % immobilization at the highest concentration of 720 mg test item/L. Thus the NOEC was determined to be 180 mg/L and the LOEC 360 mg test item/L. After 48 h the EC50 of the test item was determined to be 365 mg test item/L.

Executive summary:

The 48 –hr-acute toxicity of Ammonium formate to Daphnia magna was studied under static conditions. Test species were exposed to control, and test chemical at nominal concentration of 45.0, 90.0, 180, 360, 720 mg/L for 48 hr.  Immobilisation was observed daily.  The 48 – hour LC₅₀/EC₅₀was 365 mg/L.  The 48 –hr NOEC based on immobilization was 180 mg/L. 

Based on the results of this study, Ammonium formate does not need to be classified to Daphnia manga in accordance with Regulation (EC) No. 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals. This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age:< 24 h

Test Type (Static)

EC₅₀:365 mg/L; 95% C.l. :  307 to 434 mg/L

NOEC: 180 mg/L                      

Endpoint(s) Effected:  Immobility

Description of key information

Test according to OECD Test Guideline 202 (2002); Test Organism Age:< 24 h, Test Type (Static), EC₅₀:365 mg/L; 95% C.l. :  307 to 434 mg/L, NOEC: 180 mg/L; Endpoint(s) Effected: Immobility

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

365 mg/L